Methods and compositions relating to next generation sequencing for genetic testing in alk related cancers

ABSTRACT

Disclosed are methods, assays, and compositions for detecting the presence of a kinase inhibitor resistance. The disclosed method and primer panels work with any method for detecting nucleic acid variation in a sample including, but not limited to next generation sequencing.

BACKGROUND

Mutations of anaplastic lymphoma kinase (ALK) gene are thought to be involved in the development of subsets of numerous cancers including i) non-small cell lung carcinoma (NSCLC); ii) diffuse large B-cell lymphoma; iii) esophageal squamous cell carcinoma; iv) anaplastic large-cell lymphoma (ALCL); v) neuroblastoma (a childhood cancer that arises from the developing peripheral nervous system); and vi) the sarcomas known as inflammatory myofibroblastic tumors (IMTs). Patient outcomes with many of these malignancies are poor, due in part to the late detection of the cancers because of the lack of efficient clinical diagnostic methods. Early detection and diagnosis of ALK-mediated cancers dramatically increases survival rates within the patient population; as an example, early detection of ALK-positive anaplastic large-cell lymphoma can result in survival rates of up to 83% whereas late detection is associated in some instances with survival of only 50% of the patient population.

The critical role of deregulated ALK signaling as a driver of subsets of NSCLC, ALCL, and other ALK-dependent cancer types has been validated in clinical trials, with dramatic anti-tumor efficacy observed in response to the ALK small-molecule inhibitor crizotinib (XALKORI®, Pfizer; approved by the US FDA in August 2011). Unfortunately, despite the marked anti-tumor responses to XALKORI® seen in patients with ALK-driven tumors, most patients eventually experience progression of their cancer as a consequence of treatment resistance. For example, the median duration of progression-free survival in patients with ALK-positive NSCLC treated with Xalkori is only about 10 months. What is needed are assays the can efficiently and reliably detect kinase inhibitor-resistance mutations and therefore predict which members of a patient population is likely to develop kinase inhibitor resistance. Additionally as new generations of small-molecule inhibitors are developed, also need is a clinically applicable diagnostic test to identify resistance mutations in the ALK kinase domain and therefore to guide the rational use of these small-molecule inhibitors for the treatment of ALK-driven cancers that have lost their responsiveness to 1^(st)-generation inhibitor therapy. Moreover, once several ALK small-molecule inhibitors are approved for clinical use, optimal management of patients with ALK-driven tumors will require screening for de novo inhibitor resistance mutations by healthcare providers treating newly diagnosed patients in order to assess their inhibitor sensitivity and choose the best ALK inhibitor drug(s) for personalized therapy.

BRIEF SUMMARY

The methods, assays, and compositions disclosed herein relate to the field of detection or diagnosis of mutations that confer resistance to kinase inhibitors of a disease or condition such as cancer. In one aspect, the kinase inhibitors or ALK kinase inhibitors. Also disclosed herein are methods and assays for assessing the susceptibility or risk for developing resistance to an inhibitor, wherein the disease or condition is a cancer associated with expression of the ALK gene. It is understood and herein contemplated that the methods disclosed herein allow for rapid and sensitive detection of nucleic acid expression of mutations in ALK.

In another aspect, disclosed herein are kinase inhibitor resistance panels comprising one or more primer sets from each of the genes selected from group of genes comprising KRAS, BRAF, EGFR, ALK, and KIT.

In accordance with the purpose(s) of this invention, as embodied and broadly described herein, this invention, in one aspect, relates to an ALK kinase inhibitor resistance panel. In particular, the invention, in one aspect, relates to an ALK kinase inhibitor resistance panel comprising one or more primer sets for detecting the presence of a mutation in a gene that will confer resistance to the ALK kinase inhibitor.

Additional advantages of the disclosed methods and compositions will be set forth in part in the description which follows, and in part will be understood from the description, or may be learned by practice of the disclosed method and compositions. The advantages of the disclosed methods and compositions will be realized and attained by means of the elements and combinations particularly pointed out in the appended claims. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention as claimed.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows XALKORI®-resistance mutations identified in patient specimens. The FIGURE depicts the XALKORI®-resistance mutations in the ALK kinase domain identified to date in patient cancer specimens.

DETAILED DESCRIPTION

Before the present compounds, compositions, articles, devices, and/or methods are disclosed and described, it is to be understood that they are not limited to specific synthetic methods or specific recombinant biotechnology methods unless otherwise specified, or to particular reagents unless otherwise specified, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting.

As used in the specification and the appended claims, the singular forms “a,” “an” and “the” include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to “a pharmaceutical carrier” includes mixtures of two or more such carriers, and the like.

Ranges can be expressed herein as from “about” one particular value, and/or to “about” another particular value. When such a range is expressed, another embodiment includes from the one particular value and/or to the other particular value. Similarly, when values are expressed as approximations, by use of the antecedent “about,” it will be understood that the particular value forms another embodiment. It will be further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint, and independently of the other endpoint. It is also understood that there are a number of values disclosed herein, and that each value is also herein disclosed as “about” that particular value in addition to the value itself. For example, if the value “10” is disclosed, then “about 10” is also disclosed. It is also understood that when a value is disclosed that “less than or equal to” the value, “greater than or equal to the value” and possible ranges between values are also disclosed, as appropriately understood by the skilled artisan. For example, if the value “10” is disclosed the “less than or equal to 10” as well as “greater than or equal to 10” is also disclosed. It is also understood that the throughout the application, data is provided in a number of different formats, and that this data, represents endpoints and starting points, and ranges for any combination of the data points. For example, if a particular data point “10” and a particular data point 15 are disclosed, it is understood that greater than, greater than or equal to, less than, less than or equal to, and equal to 10 and 15 are considered disclosed as well as between 10 and 15.

In this specification and in the claims which follow, reference will be made to a number of terms which shall be defined to have the following meanings:

“Optional” or “optionally” means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

An “increase” can refer to any change that results in a larger amount of a composition or compound, such as an amplification product relative to a control. Thus, for example, an increase in the amount in amplification products can include but is not limited to a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, 250%, 300%, 350%, 400%, 450%, 500%, 550%, 600%, 700%, 800%, 900%, 1000%, 1500%, 2000%, 2500%, 3000%, 3500%, 4000%, 4500%, or 5000% increase. It is further contemplated herein that the detection an increase in expression or abundance of a DNA, mRNA, or protein relative to a control necessarily includes detection of the presence of the DNA, mRNA, or protein in situations where the DNA, mRNA, or protein is not present in the control.

“Obtaining a tissue sample” or “obtain a tissue sample” means to collect a sample of tissue either from a party having previously harvested the tissue or harvesting directly from a subject. It is understood and herein contemplated that tissue samples obtained directly from the subject can be obtained by any means known in the art including invasive and non-invasive techniques. It is also understood that methods of measurement can be direct or indirect. Examples of methods of obtaining or measuring a tissue sample can include but are not limited to tissue biopsy, tissue lavage, blood collection, aspiration, tissue swab, spinal tap, magnetic resonance imaging (MRI), Computed Tomography (CT) scan, Positron Emission Tomography (PET) scan, and X-ray (with and without contrast media). It is further understood that a “tissue” can include, but is not limited to any grouping of one or more cells or analytes to be used in a an ex vivo or in vitro assays. Such tissues include but are not limited to blood, saliva, sputum, lymph, cellular mass, and tissue collected from a biopsy.

Kinase Inhibitor Resistant Panels

In one aspect, disclosed herein are kinase inhibitor resistance panels such as, for example, an ALK kinase inhibitor panel. Kinase inhibitors are known in the art and have found use in the treatment of, amongst other things, the treatment of cancer. For example, cancers involving the overexpression or fusion of Analplastic Lymphoma Kinase can be treated through the use of a kinase inhibitor. Kinase inhibitors are known in the art and include, but are not limited to crizotinib, afatinib, Axitinib, bevacizumab, Bosutinib, Cetuximab, Dasatinib, Erlotinib, Fostamati nib, Gefitinib, Imatinib, Lapatinib, Lenvatinib, Nilotinib, Panitumumab, Pazopanib, Pegaptanib, Ranibizumab, Ruxolitinib, Sorafenib, Sunitinib, Trastuzumab, and Vemurafenib. Thus, in one aspect, disclosed herein are kinase inhibitor resistance panels for detecting susceptibility or resistance to treatment in a subject to a kinase inhibitor comprising crizotinib, afatinib, Axitinib, bevacizumab, Bosutinib, Cetuximab, Dasatinib, Erlotinib, Fostamati nib, Gefitinib, Imatinib, Lapatinib, Lenvatinib, Nilotinib, Panitumumab, Pazopanib, Pegaptanib, Ranibizumab, Ruxolitinib, Sorafenib, Sunitinib, Trastuzumab, or Vemurafenib.

Unfortunately, mutations in the ALK sequence and other genes, such as, BRAF, KIT, KRAS, and EGFR can lead to kinase inhibitor resistance. These mutations can comprise any of the mutations to ALK, KIT, BRAF, KRAS, or EGFR listed in Tables 2, 3, 4, 5, or 6. Accordingly, in a further aspect, disclosed herein are kinase inhibitor panels comprising one or more primer sets that selectively hybridize and can be used to amplify one of the genes selected from group of genes comprising KRAS (SEQ ID NO: 7718), BRAF (SEQ ID NO: 7717), EGFR (SEQ ID NO: 7716), ALK (SEQ ID NO: 7714 and SEQ ID NO: 7717 (cDNA)), and KIT. In one aspect, the kinase inhibitor resistance panel disclosed herein can comprise one or more primer set(s) that hybridizes and amplifies nucleic acid from exon 1 (SEQ ID NOs: 4601-4880 and 7181-7230) exon 2 (SEQ ID NOs: 4881-5200 and 7231-7326) or both exons 1 and 2 (SEQ ID NOs: 7327-7610) of KRAS; exon 18 (SEQ ID NOs: 1641-1760 and 5819-5934), exon 19 (SEQ ID NOs: 1761-1880), exon 20 (SEQ ID NOs: 1881-2000 and 5934-6042), exon 21 (SEQ ID NOs: 2001-2120 and 6043-6150), exon 22 (SEQ ID NOs: 2121-2240, 2321-2360, and 2401-2440), exons 18 and 19 (SEQ ID NOs: 2241-2280), exons 18, 19, and 20 (SEQ ID NOs: 6151-6274), exons 20 and 21 (SEQ ID NOs: 2281-2320 and 6275-6388), or exons 18, 19, 20, and 21 (SEQ ID NOs: 2361-2400 and 6389-6524) of EGFR; exon 8 (SEQ ID NOs: 2441-2800), exon 9 (SEQ ID NOs: 2841-3120), exon 10 (SEQ ID NOs: 3201-3360), exon 11 (SEQ ID NOs: 3361-3480), exon 12 (SEQ ID NOs: 3481-3640), exon 13 (SEQ ID NOs: 3641-3800), exon 17 (SEQ ID NOs: 4241-4600), exon 8 and 9 (SEQ ID NOs: 2801-2840), exons 9 and 10 (SEQ ID NOs: 3121-3160), exons 9, 10, and 11 (SEQ ID NOs: 3161-3200); exons 10 and 11 (SEQ ID NOs: 3801-3960), exons 12 and 13 (SEQ ID NOs: 3961-4120), or exons 10, 11, 12, and 13 (SEQ ID NOs: 4121-4240) of KIT; exons 10 and 11 (SEQ ID NOs: 6525-6832) or exons 13, 14, or 15 (SEQ ID NOs: 66833-7180) of BRAF, and/or exon 21 (SEQ ID NOs: 1-160), exon 22 (SEQ ID NOs: 401-560), exon 23 (SEQ ID NOs: 561-840 and 5311-5446), exon 24 (SEQ ID NOs: 921-1240), exon 25 (SEQ ID NOs: 1241-1600), exons 21 and 22 (SEQ ID NOs: 161-400 and 5201-5310), exons 21, 22, and 23 (SEQ ID NOs: 841-920), exons 24 and 25 (SEQ ID NOs: 1601-1640 and 5447-5576), or exons 21, 22, 23, 24, and 25 (SEQ ID NOs: 5577-5818) of ALK. As disclosed herein “primer set” refers to a forward and reverse primer pair (i.e., a left and right primer pair) that can be used together to amplify a given region of a nucleic acid (e.g., DNA, RNA, or cDNA) of interest. Thus, panels with multiple primer sets include multiple primer pairs. It is understood and herein contemplated that some primer sets may have a common forward or reverse primer and thus have an odd number of primers.

It is further understood and herein contemplated that the disclosed kinase inhibitor resistant panels can comprise a single primer sets that hybridizes to a single gene, region, or exon of a gene selected from the group of genes comprising KRAS, BRAF, EGFR, ALK, and KIT (i.e, a single primer sets for KRAS, BRAF, EGFR, ALK, or KIT); multiple primer sets that hybridize to a single gene, region, or exon of a gene selected from the group of genes comprising KRAS, BRAF, EGFR, ALK, and KIT (i.e, one or more primer sets for KRAS, BRAF, EGFR, ALK, or KIT); multiple primer sets comprising a single primer set that specifically hybridize to a single gene, region, or exon for each of the genes comprising KRAS, BRAF, EGFR, ALK, and KIT (i.e, a single primer set for each of KRAS, BRAF, EGFR, ALK, and/or KIT); or multiple primer sets comprising where in there is more than one primer set for each gene, region or exon for each of the genes selected from the group of genes comprising KRAS, BRAF, EGFR, ALK, and KIT (i.e, one or more primer sets for each of KRAS, BRAF, EGFR, ALK, and/or KIT). Thus, it is contemplated herein that the kinase inhibitor panel can comprise primer sets that recognize and specifically hybridize to a gene, region, or exon, of one or combination of the gene selected from the group consisting of KRAS, BRAF, EGFR, ALK, and KIT. For example, the panel can comprise primer sets that hybridize to a gene, region, or exon of KRAS, BRAF, EGFR, ALK, or KIT; KRAS and BRAF; KRAS and EGFR; KRAS and ALK; KRAS and KIT; BRAF and EGFR; BRAF and KIT; BRAF and ALK; EGFR and ALK; EGFR and KIT; ALK and KIT; KRAS, BRAF, and EGFR; KRAS, BRAF, and ALK; KRAS, BRAF, and KIT; KRAS, EGFR, and ALK; KRAS, EGFR, and KIT; KRAS, ALK, and KIT; BRAF, EGFR, and ALK, BRAF, EGFR, and KIT; BRAF, ALK, and KIT; EGFR, ALK, and KIT; KRAS, BRAF, EGFR, and ALK; KRAS, BRAF, EGFR, and KIT, BRAF, EGFR, ALK, and KIT; and KRAS, BRAF, EGFR, ALK, and KIT.

For example, the primer or primer sets in the kinase inhibitor resistance panel can detect any of the mutations in Tables 2-6. In another aspect, the primers or primer sets used in the inhibitor resistance panel can comprise one or more of the primers or primer sets listed in Tables 7-14 as disclosed herein and/or probes listed in Table 15 (i.e., SEQ ID NOs: 7611-7613).

Methods of Detecting the Presence of a Kinase Inhibitor Resistant Cancer

The disclosed kinase inhibitor resistant panels, in one aspect, contain primers or primer sets for the detection of mutations that confer kinase inhibitor resistance. Thus, in another aspect disclosed herein are methods and assays for the detection of kinase inhibitor resistant forms of an ALK-related cancer. For example, disclosed herein are methods and assays for the detection of kinase inhibitor resistance, such as, for example ALK kinase inhibitor resistance, comprising obtaining a tissue sample from a subject with a cancer, such as a kinase related cancer (e.g., ALK-related cancers); conducting a high throughput sequencing (also known as next generation sequencing) reaction on the sample, wherein the presence of a mutation in the nucleic acid sequence of a gene associated with kinase inhibitor resistance indicates that that the cancer is resistant or will become resistant to a kinase inhibitor. In one aspect, the mutation can be a nucleic acid mutation in ALK, EGFR, KRAS, BRAF, or KIT. For example, the mutation can be any mutation listed in Tables 2-6. In a further aspect, the disclosed methods and assays for detection of kinase inhibitor resistance can comprise performing next generation sequencing using a kinase inhibitor resistant panel as disclosed herein which comprises a primer or primer set that hybridizes and amplifies nucleic acid from exon 1 or 2 of KRAS; exon 18, 19, 20, 21 or 22 of EGFR; exon 8, 9, 10, 11, 12, 13, or 17 of KIT; exon 10, 11, 13, 14, or 15 of BRAF, and/or exon 21, 22, 23, 24, or 25 of ALK. For example, the primer or primer set can comprise any of the primers or primer sets disclosed in Tables 7-14. Thus, disclosed herein are methods wherein the one or more primer set(s) that hybridizes and amplifies nucleic acid from exon 1 (SEQ ID NOs: 4601-4880 and 7181-7230) exon 2 (SEQ ID NOs: 4881-5200 and 7231-7326) or both exons 1 and 2 (SEQ ID NOs: 7327-7610) of KRAS; exon 18 (SEQ ID NOs: 1641-1760 and 5819-5934), exon 19 (SEQ ID NOs: 1761-1880), exon 20 (SEQ ID NOs: 1881-2000 and 5934-6042), exon 21 (SEQ ID NOs: 2001-2120 and 6043-6150), exon 22 (SEQ ID NOs: 2121-2240, 2321-2360, and 2401-2440), exons 18 and 19 (SEQ ID NOs: 2241-2280), exons 18, 19, and 20 (SEQ ID NOs: 6151-6274), exons 20 and 21 (SEQ ID NOs: 2281-2320 and 6275-6388), or exons 18, 19, 20, and 21 (SEQ ID NOs: 2361-2400 and 6389-6524) of EGFR; exon 8 (SEQ ID NOs: 2441-2800), exon 9 (SEQ ID NOs: 2841-3120), exon 10 (SEQ ID NOs: 3201-3360), exon 11 (SEQ ID NOs: 3361-3480), exon 12 (SEQ ID NOs: 3481-3640), exon 13 (SEQ ID NOs: 3641-3800), exon 17 (SEQ ID NOs: 4241-4600), exon 8 and 9 (SEQ ID NOs: 2801-2840), exons 9 and 10 (SEQ ID NOs: 3121-3160), exons 9, 10, and 11 (SEQ ID NOs: 3161-3200); exons 10 and 11 (SEQ ID NOs: 3801-3960), exons 12 and 13 (SEQ ID NOs: 3961-4120), or exons 10, 11, 12, and 13 (SEQ ID NOs: 4121-4240) of KIT; exons 10 and 11 (SEQ ID NOs: 6525-6832) or exons 13, 14, or 15 (SEQ ID NOs: 66833-7180) of BRAF, and/or exon 21 (SEQ ID NOs: 1-160), exon 22 (SEQ ID NOs: 401-560), exon 23 (SEQ ID NOs: 561-840 and 5311-5446), exon 24 (SEQ ID NOs: 921-1240), exon 25 (SEQ ID NOs: 1241-1600), exons 21 and 22 (SEQ ID NOs: 161-400 and 5201-5310), exons 21, 22, and 23 (SEQ ID NOs: 841-920), exons 24 and 25 (SEQ ID NOs: 1601-1640 and 5447-5576), or exons 21, 22, 23, 24, and 25 (SEQ ID NOs: 5577-5818) of ALK.

It is understood that the disclosed methods can further comprise synthesizing cDNA from the nucleic acid extracted from a tissue sample before detection of a mutation in ALK, EGFR, KRAS, BRAF, or KIT. Thus, in one aspect, disclosed herein are methods for detecting kinase inhibitor resistance in a cancer in a subject, for example ALK kinase inhibitor resistance, comprising obtaining a tissue sample from a subject with a cancer, such as a kinase related cancer (e.g., ALK-related cancers); synthesixing cDNA from the tissue sample, and conducting a high throughput sequencing (also known as next generation sequencing) reaction on the sample, wherein the presence of a mutation in the nucleic acid sequence of a gene associated with kinase inhibitor resistance indicates that that the cancer is resistant or will become resistant to a kinase inhibitor.

It is further understood and herein contemplated that the subject of the disclosed methods can be a subject that has been previously diagnosed with a cancer including but not limited to inflammatory breast cancer, non-small cell lung carcinoma, esophageal squamous cell carcinoma, colorectal carcinoma, Inflammatory myofibroblastic tumor, familial and sporadic neuroblastoma. In yet another aspect, the subject has been previously diagnosed with a cancer that results from ALK, ROS1, RET, DEPDC1 overexpression, dysregulation, or fusion. Examples of such fusions include but are not limited to nucleophosmin-ALK (NPM-ALK), 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase/IMP cyclohydrolase (ATIC-ALK), clathrin heavy chain-ALK (CLTC-ALK), kinesin-1 heavy chain gene-ALK (KIF5B-ALK); Ran-binding protein 2-ALK (RANBP2-ALK), SEC31L1-ALK, tropomyosin-3-ALK (TPM3-ALK), tropomyosin-4-ALK (TPM4-ALK), TRK-fused gene (Large)-ALK (TFG_(L)-ALK), TRK-fused gene (Small)-ALK (TFG_(S)-ALK), CARS-ALK, EML4-ALK, 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase/IMP cyclohydrolase-ALK (ATIC-ALK), ALO17-ALK, moesin-ALK (MSN-ALK), non-muscle myosin heavy chain gene-ALK (MYH9-ALK), and TRK-fused gene (Extra Large)-ALK (TFGxL-ALK). In a further aspect, the present methods could not only be used to diagnose a kinase inhibitor resistant cancer, but diagnose the cancer itself as the subject with a kinase inhibitor resistant cancer would necessarily not only have a cancer, but have a kinase related cancer such as those disclosed herein.

Therefore, in one aspect, disclosed herein are methods for the detection of kinase inhibitor resistance comprising obtaining a tissue sample from a subject with a cancer and conducting a high throughput sequencing (also known as next generation sequencing) reaction on the sample using one or more primer sets or primer panels with primer sets that specifically hybridizes to one or more of the genes selected from the group consisting of ALK, KRAS, EGFR, KIT, and BRAF, wherein the presence of a mutation in the nucleic acid sequence of a gene associated with kinase inhibitor resistance indicates that that the cancer is resistant or will become resistant to a kinase inhibitor.

Also disclosed are methods, wherein at least one primer sets hybridizes and amplifies nucleic acid from exon 1 or 2 of KRAS, wherein at least one primer sets hybridizes and amplifies nucleic acid from exon 18, 19, 20, 21 or 22 of EGFR, wherein at least one primer sets hybridizes and amplifies nucleic acid from exon 21, 22, 23, 24, or 25 of ALK, wherein at least one primer sets hybridizes and amplifies nucleic acid from exon 8, 9, 10, 11, 12, 13, or 17 of KIT, and/or wherein at least one primer sets hybridizes and amplifies nucleic acid from exon 10, 11, 13, 14, or 15 of BRAF.

In one aspect, disclosed are methods, wherein one or more KRAS hybridizing primers or primer sets comprise one or more of the primers of Tables 10 and/or 14 (SEQ ID NOs: 4601-5200 and 7181-7610); wherein one or more EGFR hybridizing primers or primer sets comprise one or more of the primers of Tables 8 and/or 12 (1641-2440 and 5819-6524); wherein one or more ALK hybridizing primers or primer sets comprise one or more of the primers of Tables 7 and/or 11 (SEQ ID NOs: 1-1640 and 5201-5818); wherein one or more KIT hybridizing primers or primer sets comprise one or more of the primers of Table 9 (SEQ ID NOs: 2441-4600); and/or wherein one or more BRAF hybridizing primers or primer sets comprise one or more of the primers of Table 13 (SEQ ID NOs: 6525-7180).

In one aspect are methods comprising the use of a kinase inhibitor resistance panel, wherein the panel comprises 2, 3, 4, 5, 6, 7, 8, 9, 10, or more primer sets for one or more of the genes selected from group of genes comprising KRAS, BRAF, EGFR, ALK, and KIT.

In another aspect, disclosed are methods wherein the panel comprises one or more primer sets for 2, 3, 4, of all 5 of the genes selected from group of genes comprising KRAS, BRAF, EGFR, ALK, and KIT.

Also disclosed are methods, wherein the kinase inhibitor is selected from the group consisting of crizotinib, afatinib, Axitinib, bevacizumab, Bosutinib, Cetuximab, Dasatinib, Erlotinib, Fostamati nib, Gefitinib, Imatinib, Lapatinib, Lenvatinib, Nilotinib, Panitumumab, Pazopanib, Pegaptanib, Ranibizumab, Ruxolitinib, Sorafenib, Sunitinib, Trastuzumab, and Vemurafenib.

Methods, Assays, and Primer Panels for Assessing the Suitability of ALK Directed Treatments

Though not wishing to be bound by current theories, it is believed that inhibition of these over-expression or aberrant expressions of ALK with small-molecule drug candidates abrogates related abnormal cell proliferation and promotes apoptosis in ALK-related tumor cell lines. Furthermore, both preclinical animal models and the early clinical experience with these inhibitors indicate that ALK small-molecule inhibitors not only possess marked antitumor activity against ALK-related cancers but are also very well tolerated with no limiting target-associated toxicities. Therefore, such small molecules can be used to treat ALK-driven cancers.

However, the presence of a mutation in one of the genes associated with an ALK-related cancer can confer resistance to treatment with a kinase inhibitor, such as an ALK kinase inhibitor. Nevertheless, knowledge of the presence of said mutation can still be useful to the practicing physician in assessing the suitability of a treatment or prescribing a particular treatment regimen. For example, the presence of a mutation in a gene which confers kinase inhibitor resistance, such as, for example, ALK kinase inhibitor resistance, can inform the skilled artisan to choose a particular kinase inhibitor over another due to the presence of a mutation affecting one kinase inhibitor and not the other. Alternatively, the presence of a mutation can inform the physician to discontinue the course of treatment with one kinase inhibitor due to detection of kinase inhibitor resistance and select a different kinase inhibitor to which the patient is not yet resistant. Accordingly, disclosed herein are methods and assays for assessing the suitability of an ALK inhibitor treatment for a cancer, for example, NSCLC, in a subject comprising performing high throughput sequencing on nucleic acid from a tissue sample from the subject; wherein the presence of a mutation in ALK, EGFR, BRAF, KRAS, or KIT indicates a cancer that comprises resistance to an ALK kinase inhibitor. In one aspect, disclosed herein are methods and assays for assessing a subject's suitability for treatment with a kinase inhibitor comprising obtaining a tissue sample from a subject with a cancer, such as a kinase related cancer (e.g., ALK-related cancers); detecting the presence of a mutation through sequencing or other nucleic acid detection technique for the presence of a mutation in the nucleic acid sequence of a gene associated with kinase inhibitor resistance indicates that that the cancer is resistant or will become resistant to a kinase inhibitor and therefore continued use of an inhibitor to which the cancer has become resistant or to which the cancer is already resistant should be discontinued in favor of a cancer to which resistance has not developed.

It is understood and herein contemplated that any of the disclosed nucleic acid sequencing techniques disclosed herein can be used in these methods. Thus, disclosed herein are methods and assays assessing the suitability of an ALK kinase inhibitor treatment for an ALK related cancer in a subject comprising conducting high throughput sequencing (also known as next generation sequencing) on nucleic acid such as mRNA or DNA from a tissue sample from the subject; wherein the sequencing reaction reveals the nucleic acid sequence for one or more exons of KIT, BRAF, KRAS, EGFR, and ALK; and wherein the presence of one or more mutations in KIT, BRAF, KRAS, EGFR, and/or ALK indicates the presence of kinase inhibitor resistance. The mutations can occur in any exon of KIT, BRAF, KRAS, EGFR, and ALK. Thus, for example, the mutations can occur in and therefore the primers or primer sets can hybridize to exon 1 or 2 of KRAS; exon 18, 19, 20, 21 r 22 of EGFR; exon 8, 9, 10, 11, 12, 13, or 17 of KIT; exon 10, 11, 13, 14, or 15 of BRAF, and/or exon 21, 22, 23, 24, or 25 of ALK. In one aspect, the mutation can comprise any one or more of the mutations listed in Tables 2-6. It is further understood that the disclosed methods and assays can further comprise any of the primers disclosed herein in Tables 7-14 or probes listed in Table 15 and utilize the multiplexing PCR techniques disclosed.

In another aspect, two or more of the disclosed primers and primer sets can comprise a primer panel can be used in methods and assays for the assessment of the suitability of a kinase inhibitor for the treatment of a subjects' cancer. In one aspect, the primer panel comprises one or more primers that can detect a nucleic acid mutation in ALK, BRAF, EGFR, KRAS, or KIT. In a further aspect, the primers or primer sets that hybridizes and amplifies nucleic acid from exon 1 or 2 of KRAS; exon 18, 19, 20, 21 or 22 of EGFR; exon 8, 9, 10, 11, 12, 13, or 17 of KIT; exon 10, 11, 13, 14, or 15 of BRAF, and/or exon 21, 22, 23, 24, or 25 of ALK. In another aspect, the disclosed primer panel can comprise any primer or primer set which detects one or more of the mutations found in Tables 2-6. For example, the primer or primer set can comprise any of the primers or primer sets disclosed in Tables 7-14.

In another aspect, knowledge of kinase inhibitor resistant cancer can be used to screen for a drug that is not a kinase inhibitor. Thus, in one aspect, disclosed herein are methods of screening for a drug to treat a subject with a cancer comprising obtaining a tissue sample from a subject with a cancer, such as a kinase related cancer (e.g., ALK-related cancers); conducting a high throughput sequencing (also known as next generation sequencing) reaction on the sample, wherein the presence of a mutation in the nucleic acid sequence of a gene, region, or exon associated with kinase inhibitor resistance indicates that that the subject has a cancer is resistant or will become resistant to a kinase inhibitor, and contacting a tissue sample from subject with a cancer with an agent; wherein an agent that inhibits or reduces the growth or development of a kinase inhibitor resistant cancer is not a kinase inhibitor. The disclosed methods can further comprise the sue of the kinase inhibitor resistant panels disclosed herein or any of the primers, primer sets or probes disclosed herein. The methods can also further comprise the treatment of a subject with a kinase inhibitor resistant cancer with an agent that is identified in the method as not being a kinase inhibitor or discontinuing treatment in a subject with kinase inhibitor resistant cancer with an agent that has been found to be a kinase inhibitor.

Methods of Identifying Subjects for Participation in Clinical Trials to Screen for New Cancer Treatments.

In one aspect, it is contemplated herein that the identification of individuals with a kinase inhibitor resistant cancer can be useful for establishing clinical trials to screen for drugs that can be used to treat individuals with kinase inhibitor resistant cancers. Thus, in one aspect, disclosed herein are methods for identifying a subject for screening for a drug that can treat a cancer in a subject with a kinase inhibitor resistant cancer, for example ALK kinase inhibitor resistance, comprising obtaining a tissue sample from a subject with a cancer, such as a kinase related cancer (e.g., ALK-related cancers); and conducting a high throughput sequencing (also known as next generation sequencing) reaction on the sample, wherein the presence of a mutation in the nucleic acid sequence of a gene, region, or exon associated with kinase inhibitor resistance indicates that that the subject has a cancer is resistant or will become resistant to a kinase inhibitor and the subject can be used in trials to screen for a drug to which a kinase inhibitor resistant subject will respond. In one aspect, the mutation can be a nucleic acid mutation in ALK, EGFR, KRAS, BRAF, or KIT. For example, the mutation can be any mutation listed in Tables 2-6. In one aspect, said methods can further comprise synthesizing cDNA from the tissue sample of the subject.

It is understood and herein contemplated that the disclosed methods can be used in conjunction with any of the kinase inhibitor resistant panels, primer sets, or probes disclosed herein. For example, the disclosed methods can be performed using a primer or primer set that hybridizes and amplifies nucleic acid from exon 1 or 2 of KRAS; exon 18, 19, 20, 21 or 22 of EGFR; exon 8, 9, 10, 11, 12, 13, or 17 of KIT; exon 10, 11, 13, 14, or 15 of BRAF, and/or exon 21, 22, 23, 24, or 25 of ALK. For example, the primer or primer set can comprise any of the primers or primer sets disclosed in Tables 7-14. Thus, disclosed herein are methods wherein the one or more primer set(s) that hybridizes and amplifies nucleic acid from exon 1 (SEQ ID NOs: 4601-4880 and 7181-7230) exon 2 (SEQ ID NOs: 4881-5200 and 7231-7326) or both exons 1 and 2 (SEQ ID NOs: 7327-7610) of KRAS; exon 18 (SEQ ID NOs: 1641-1760 and 5819-5934), exon 19 (SEQ ID NOs: 1761-1880), exon 20 (SEQ ID NOs: 1881-2000 and 5934-6042), exon 21 (SEQ ID NOs: 2001-2120 and 6043-6150), exon 22 (SEQ ID NOs: 2121-2240, 2321-2360, and 2401-2440), exons 18 and 19 (SEQ ID NOs: 2241-2280), exons 18, 19, and 20 (SEQ ID NOs: 6151-6274), exons 20 and 21 (SEQ ID NOs: 2281-2320 and 6275-6388), or exons 18, 19, 20, and 21 (SEQ ID NOs: 2361-2400 and 6389-6524) of EGFR; exon 8 (SEQ ID NOs: 2441-2800), exon 9 (SEQ ID NOs: 2841-3120), exon 10 (SEQ ID NOs: 3201-3360), exon 11 (SEQ ID NOs: 3361-3480), exon 12 (SEQ ID NOs: 3481-3640), exon 13 (SEQ ID NOs: 3641-3800), exon 17 (SEQ ID NOs: 4241-4600), exon 8 and 9 (SEQ ID NOs: 2801-2840), exons 9 and 10 (SEQ ID NOs: 3121-3160), exons 9, 10, and 11 (SEQ ID NOs: 3161-3200); exons 10 and 11 (SEQ ID NOs: 3801-3960), exons 12 and 13 (SEQ ID NOs: 3961-4120), or exons 10, 11, 12, and 13 (SEQ ID NOs: 4121-4240) of KIT; exons 10 and 11 (SEQ ID NOs: 6525-6832) or exons 13, 14, or 15 (SEQ ID NOs: 66833-7180) of BRAF, and/or exon 21 (SEQ ID NOs: 1-160), exon 22 (SEQ ID NOs: 401-560), exon 23 (SEQ ID NOs: 561-840 and 5311-5446), exon 24 (SEQ ID NOs: 921-1240), exon 25 (SEQ ID NOs: 1241-1600), exons 21 and 22 (SEQ ID NOs: 161-400 and 5201-5310), exons 21, 22, and 23 (SEQ ID NOs: 841-920), exons 24 and 25 (SEQ ID NOs: 1601-1640 and 5447-5576), or exons 21, 22, 23, 24, and 25 (SEQ ID NOs: 5577-5818) of ALK.

Methods of Detecting a Kinase Inhibitor Resistance in an ALK-Related Cancer

In another aspect, the disclosed methods and assays relate to the detection or diagnosis of the presence of a kinase inhibitor resistance, such as, for example, ALK kinase inhibitor resistance, in a disease or condition such as a cancer and methods and assays for the determination of susceptibility or resistance to therapeutic treatment for a disease or condition such as a cancer in a subject comprising detecting the presence or measuring the expression level of nucleic acid (for example, DNA, mRNA, cDNA, RNA, etc) through the use of next generation sequencing (NGS) from a tissue sample from the subject; wherein the presence of a mutations in the nucleic acid code of the KIT, BRAF, KRAS, EGFR, or ALK gene or the ALK gene portion of an ALK fusion construct indicates the presence of a cancer that is resistant to a kinase inhibitor. In one aspect, the cancer is associated with amplification, overexpression, nucleic acid variation, truncation, or gene fusion of ALK. It is understood, that the kinase inhibitor resistance panels disclosed herein can be used to perform said methods and the detection of one or more of the mutations in Tables 2-6 indicates the presence of kinase inhibitor resistance. In one aspect, the disclosed methods can further comprise discontinuing use of a kinase inhibitor to treat a cancer in a subject that has been identified with a kinase inhibitor resistant cancer. In another embodiment, the disclosed methods can further comprise treating a subject with a kinase inhibitor resistant cancer with a chemotherapeutic that is not a kinase inhibitor. Thus, in one aspect, disclosed herein are methods of treating a subject with a kinase inhibitor resistant cancer (such as, for example, an ALK kinase inhibitor resistant cancer) comprising obtaining a tissue sample from a subject with a cancer, such as a kinase related cancer (e.g., ALK-related cancers); conducting a high throughput sequencing (also known as next generation sequencing) reaction on the sample, wherein the presence of a mutation in the nucleic acid sequence of a gene, region, or exon associated with kinase inhibitor resistance indicates that that the subject has a cancer is resistant or will become resistant to a kinase inhibitor; and treating the subject with a chemotherapeutic that is not a kinase inhibitor. Also disclosed are methods of treating a subject without a kinase inhibitor resistant cancer comprising obtaining a tissue sample from a subject with a cancer, such as a kinase related cancer (e.g., ALK-related cancers); conducting a high throughput sequencing (also known as next generation sequencing) reaction on the sample, wherein the absence of a mutation in the nucleic acid sequence of a gene, region, or exon associated with kinase inhibitor resistance indicates that that the subject does not have a cancer is resistant nor will become resistant to a kinase inhibitor; and treating the subject with a kinase inhibitor.

Anaplastic Lymphoma Kinase (ALK)

ALK (SEQ ID NO: 7714 (Genbank Accession No. U62540 (human coding sequence)) is a receptor tyrosine kinase (RTK) of the insulin receptor superfamily encoded by the ALK gene and is normally expressed primarily in the central and peripheral nervous systems. The 1620aa ALK polypeptide comprises a 1030aa extracellular domain which includes a 26aa amino-terminal signal peptide sequence, and binding sites located between residues 391 and 401 for the ALK ligands pleiotrophin (PTN) and midkine (MK). Additionally, the ALK polypeptide comprises a kinase domain (residues 1116-1383) which includes three tyrosines responsible for autophosphorylation within the activation loop at residues 1278, 1282, and 1283. ALK amplification, overexpression, and mutations have been shown to constitutively activate the kinase catalytic function of the ALK protein, with the deregulated mutant ALK in turn activating downstream cellular signaling proteins in pathways that promote aberrant cell proliferation. In fact, the mutations that result in dysregulated ALK kinase activity are associated with several types of cancers.

ALK fusions represent the most common mutation of this tyrosine kinase. Such fusions include but are not limited to nucleophosmin-ALK (NPM-ALK), 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase/IMP cyclohydrolase (ATIC-ALK), clathrin heavy chain-ALK (CLTC-ALK), kinesin-1 heavy chain gene-ALK (KIF5B-ALK); Ran-binding protein 2-ALK (RANBP2-ALK), SEC31L1-ALK, tropomyosin-3-ALK (TPM3-ALK), tropomyosin-4-ALK (TPM4-ALK), TRK-fused gene (Large)-ALK (TFG_(L)-ALK), TRK-fused gene (Small)-ALK (TFGs-ALK), CARS-ALK, EML4-ALK, 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase/IMP cyclohydrolase-ALK (ATIC-ALK), ALO17-ALK, moesin-ALK (MSN-ALK), non-muscle myosin heavy chain gene-ALK (MYH9-ALK), and TRK-fused gene (Extra Large)-ALK (TFGxL-ALK). Six ALK fusions, CARS-ALK. CLTC-ALK, RANBP2-ALK, SEC31L1-ALK, TPM3-ALK, and TPM4-ALK have been identified in IMTs. TPM3-ALK, TPM4-ALK and CLTC-ALK fusions have been detected in both classical T- or null-cell lymphomas and IMT sarcomas, whereas CARS-ALK, RANBP2-ALK, and SEC31L1-ALK occur in IMT. CLTC-ALK and NPM-ALK also occur in B-cell plasmablastic/immunoblastic lymphomas. The TPM4-ALK fusion occurs in esophageal squamous cell carcinomas, and the ALK fusion EML4-ALK, TFG-ALK and KIF5B-ALK are found in non-small cell lung cancers. EML4-ALK has also recently been identified in both colorectal and breast carcinomas as well.

ALK fusions are associated with several known cancer types. It is understood that one or more ALK fusions can be associated with a particular cancer. It is further understood that there are several types of cancer associated with ALK fusions including but not limited to anaplastic large-cell lymphoma (ALCL), neuroblastoma, breast cancer, ovarian cancer, colorectal carcinoma, non-small cell lung carcinoma, diffuse large B-cell lymphoma, esophageal squamous cell carcinoma, anaplastic large-cell lymphoma, neuroblastoma, inflammatory myofibroblastic tumors, malignant histiocytosis, and glioblastomas.

ALCL. anaplastic large-cell lymphomas comprise ˜2.5% of all NHL; within the pediatric age group specifically, ˜13% of all NHL (30-40% of all childhood large-cell lymphomas) are of this type. Studies of ALCL patients now divide this NHL into ALK-positive and ALK-negative subsets; ˜60% of all ALCLs are caused by ALK fusions. For unclear reasons, ALK-positive ALCL patients fare significantly better following CHOP based multi-agent conventional chemotherapy than those with ALK-negative disease (with overall 5-year survivals of ˜75% vs. ˜35%, respectively). However, more than a third of patients suffer multiple relapses following chemotherapy, thus the 5-year disease-free survival of ALK-positive ALCL is only ˜40%.

ALK+ Diffuse large B-cell lymphoma. In 2003, ALK fusions were shown to occur in a non-ALCL form of NHL with the description of CLTC-ALK or NPM-ALK in diffuse large B-cell lymphomas (ALK+ DLBCLs). Consistent with their B-lineage, these NHLs express cytoplasmic IgA and plasma cell markers, and possess an immunoblastic morphology. Translational research studies revealed the t(2; 17) and CLTC-ALK mRNA in the majority of these lymphomas, while immunolabeling confirmed granular ALK staining identical to that observed in CLTC-ALK-positive ALCL. As for all other ALK fusion partner proteins, a self-association motif in the CLTC portion of CLTC-ALK mediates constitutive self-association and activation of the fusion kinase to drive lymphomagenesis. ALK+ DLBCLs occur predominately in adults; however, the t(2; 5) and NPM-ALK mRNA in pediatric lymphomas are phenotypically identical to CLTC-ALK-positive adult B-NHLs. Approximately 0.5-1% of all DLBCL is thought to be ALK-positive. The identification of DLBCLs caused by mutant ALK is important because patients with these lymphomas have outcomes that are much inferior to ALK-negative DLBCL patients following CHOP-based treatments; thus, ALK+ DLBCL patients should strongly be considered as candidates for ALK-targeted kinase inhibitor therapy.

ALK+ systemic histiocytosis. ALK fusions were described in 2008 in another hematopoietic neoplasm, systemic histiocytosis. Three cases of this previously uncharacterized form of histiocytosis, which presents in early infancy, exhibited ALK immunoreactivity and the one case analyzed molecularly expressed TPM3-ALK.

In addition to the aforementioned hematological malignancies in which constitutively activated ALK fusions have been shown to be a causative mechanism in many cases, the genesis of subsets of various solid tumors in some instances, very common human tumors such as non-small cell lung cancer, colorectal and breast cancers has recently been demonstrated to be due to aberrantly activated ALK.

Inflammatory myofibroblastic tumor. The first non-hematopoietic tumor discovered to express ALK fusions was the sarcoma known as inflammatory myofibroblastic tumor (IMT), a spindle cell proliferation in the soft tissue and viscera of children and young adults (mean age at diagnosis ˜10 years). Many IMTs are indolent and can be cured by resection. However, locally recurrent, invasive, and metastatic IMTs are not uncommon and current chemo- and radio-therapies are completely ineffective. Disclosed herein is the involvement of chromosome 2p23 (the location of the ALK gene) in IMTs, as well as ALK gene rearrangement. ALK immunoreactivity in 7 of 11 IMTs has been shown and TPM3-ALK and TPM4-ALK were identified in several cases. Additionally, two additional ALK fusions in IMT, CLTC- and RanBP2-ALK were identified. ALK fusions have also been examined by immunostaining in 73 IMTs, finding 60% (44 of the 73 cases) to be ALK-positive. Thus, ALK deregulation is of pathogenic importance in a majority of IMTs.

Non-small cell lung carcinoma. The role of ALK fusions in cancer expanded further with the description of the novel EML4-ALK chimeric protein in 5 of 75 (6.7%) Japanese non-small cell lung carcinoma patients. Shortly thereafter, the existence of ALK fusions in lung cancer was corroborated by a different group who found 6 of 137 (4.4%) Chinese lung cancer patients to express ALK fusions (EML4-ALK, 3 pts; TFG-ALK, 1 pt; X-ALK. Two common themes have emerged—1) ALK fusions occur predominately in patients with adenocarcinoma (although occasional ALK-positive NSCLCs of squamous or mixed histologies are observed), mostly in individuals with minimal/no smoking history, and 2) ALK abnormalities usually occur exclusive of other common genetic abnormalities (e.g., EGFR and KRAS mutations). The exact percentage of NSCLCs caused by ALK fusions is not yet clear but estimates based on reports in the biomedical literature suggest a range of ˜5-10%.

Esophageal squamous cell carcinoma. In 45 Iranian patients, a proteomics approach identified proteins under or over-represented in esophageal squamous cell carcinomas (ESCCs); TPM4-ALK was among those proteins over-represented. A second proteomics-based ESCC study—in this case, in Chinese patients—identified TPM4-ALK in these tumors as well.

Colorectal carcinoma, breast cancer. Three human tumor types—colorectal, breast, and non-small cell lung cancers were surveyed for the presence of the EML4-ALK fusion (other ALK mutations were not assessed in this study). In addition to confirming the expression of EML4-ALK in NSCLC (in 12 of 106 specimens studied, 11.3%), a subsets of breast (5 of 209 cases, 2.4%) and colorectal (2 of 83 cases, 2.4%) carcinomas were EML4-ALK-positive. In addition to known EML4-ALK variants 1 (E13; A20) and 2 (E20; A20), a novel variant (E21; A20) was found in colorectal carcinoma.

ALK in familial and sporadic neuroblastoma. Neuroblastoma is the most common extracranial solid tumor of childhood, and is derived from the developing neural crest. A small subset (˜1-2%) of neuroblastomas exhibit a familial predisposition with an autosomal dominant inheritance. Most neuroblastoma patients have aggressive disease associated with survival probabilities <40% despite intensive chemo- and radio-therapy, and the disease accounts for ˜15% of all childhood cancer mortality. ALK had previously been found to be constitutively activated also due to high-level over-expression as a result of gene amplification in a small number of neuroblastoma cell lines, in fact, ALK amplification occurs in ˜15% of neuroblastomas in addition to activating point mutations. These missense mutations in ALK have been confirmed as activating mutations that drive neuroblastoma growth; furthermore, incubation of neuroblastoma cell lines with ALK small-molecule inhibitors reveal those cells with ALK activation (but not cell lines with normal levels of expression of wild-type ALK) to exhibit robust cytotoxic responses.

The sensitive detection of a mutation at a known site in DNA is readily done with existing technologies. Allele specific primers can be designed to target a mutation at a known location such that its signal can be preferentially amplified over wild-type DNA.

Next Generation Sequencing for Genetic Testing

From a technical perspective High-throughput or Next Generation Sequencing (NGS) represents an attractive option for detecting the somatic mutations within a gene. Unlike PCR, microarrays, high-resolution melting and mass spectrometry, which all indirectly infer sequence content, NGS directly ascertains the identity of each base and the order in which they fall within a gene. The newest platforms on the market have the capacity to cover an exonic region 10,000 times over, meaning the content of each base position in the sequence is measured thousands of different times. This high level of coverage ensures that the consensus sequence is extremely accurate and enables the detection of rare variants within a heterogeneous sample. For example, in a sample extracted from FFPE tissue, relevant mutations are only present at a frequency of 1% with the wild-type allele comprising the remainder. When this sample is sequenced at 10,000× coverage, then even the rare allele, comprising only 1% of the sample, is uniquely measured 100 times over. Thus, NGS can provide reliably accurate results with very high sensitivity, making it ideal for clinical diagnostic testing of FFPEs and other mixed samples.

In one aspect, disclosed herein are methods and assays for detecting kinase inhibitor resistance or determining the susceptibility to a particular kinase inhibitor treatment in an ALK-related cancer comprising performing next generation sequencing on a tissue sample obtained from a subject with an ALK-related cancer, wherein the presence of a nucleic acid variation in the ALK, BRAF, EGFR, KIT, or KRAS sequence of the tissue sample at a nucleic acid residue indicates that presence of kinase inhibitor resistance. For example, the methods and assays for detecting kinase inhibitor resistance or determining the susceptibility or developing kinase inhibitor resistance in an ALK-related cancer or determining the suitability of a particular kinase inhibitor for use in treating an ALK-related cancer in a subject can comprise the detection of any of the mutations in Tables 2-6. It is understood that the methods and assays can further comprise comparing the sequence to known kinase inhibitor resistance mutations list and determining what if any kinase inhibitors are affected by the mutation and altering or maintaining treatment as appropriate to utilize kinase inhibitors that are unaffected by the mutation. As the disclosed methods and assays employ the use of primers or primer sets to detect mutations that confer kinase inhibitor resistance, also disclosed herein are primer panels for use in next generation sequencing for the determination of kinase inhibitor resistance comprising one or more primer sets from each of KIT, BRAF, KRAS, EGFR, and ALK, for example, the disclosed primer panels, methods, and assays can comprise one or more of the primers or primer sets listed in Tables 7-14.

Examples of Next Generation Sequencing techniques include, but are not limited to Massively Parallel Signature Sequencing (MPSS), Polony sequencing, pyrosequencing, Reversible dye-terminator sequencing, SOLiD sequencing, Ion semiconductor sequencing, DNA nanoball sequencing, Helioscope single molecule sequencing, Single molecule real time (SMRT) sequencing, Single molecule real time (RNAP) sequencing, and Nanopore DNA sequencing.

MPSS was a bead-based method that used a complex approach of adapter ligation followed by adapter decoding, reading the sequence in increments of four nucleotides; this method made it susceptible to sequence-specific bias or loss of specific sequences.

Polony sequencing, combined an in vitro paired-tag library with emulsion PCR, an automated microscope, and ligation-based sequencing chemistry to sequence an E. coli genome at an accuracy of >99.9999% and a cost approximately 1/10 that of Sanger sequencing.

A parallelized version of pyrosequencing, the method amplifies DNA inside water droplets in an oil solution (emulsion PCR), with each droplet containing a single DNA template attached to a single primer-coated bead that then forms a clonal colony. The sequencing machine contains many picolitre-volume wells each containing a single bead and sequencing enzymes. Pyrosequencing uses luciferase to generate light for detection of the individual nucleotides added to the nascent DNA, and the combined data are used to generate sequence read-outs. This technology provides intermediate read length and price per base compared to Sanger sequencing on one end and Solexa and SOLiD on the other.

A sequencing technology based on reversible dye-terminators. DNA molecules are first attached to primers on a slide and amplified so that local clonal colonies are formed. Four types of reversible terminator bases (RT-bases) are added, and non-incorporated nucleotides are washed away. Unlike pyrosequencing, the DNA can only be extended one nucleotide at a time. A camera takes images of the fluorescently labeled nucleotides, then the dye along with the terminal 3′ blocker is chemically removed from the DNA, allowing the next cycle.

SOLiD technology employs sequencing by ligation. Here, a pool of all possible oligonucleotides of a fixed length are labeled according to the sequenced position. Oligonucleotides are annealed and ligated; the preferential ligation by DNA ligase for matching sequences results in a signal informative of the nucleotide at that position. Before sequencing, the DNA is amplified by emulsion PCR. The resulting bead, each containing only copies of the same DNA molecule, are deposited on a glass slide. The result is sequences of quantities and lengths comparable to Illumina sequencing.

Ion semiconductor sequencing is based on using standard sequencing chemistry, but with a novel, semiconductor based detection system. This method of sequencing is based on the detection of hydrogen ions that are released during the polymerization of DNA, as opposed to the optical methods used in other sequencing systems. A microwell containing a template DNA strand to be sequenced is flooded with a single type of nucleotide. If the introduced nucleotide is complementary to the leading template nucleotide it is incorporated into the growing complementary strand. This causes the release of a hydrogen ion that triggers a hypersensitive ion sensor, which indicates that a reaction has occurred. If homopolymer repeats are present in the template sequence multiple nucleotides will be incorporated in a single cycle. This leads to a corresponding number of released hydrogens and a proportionally higher electronic signal.

DNA nanoball sequencing is a type of high throughput sequencing technology used to determine the entire genomic sequence of an organism. The method uses rolling circle replication to amplify small fragments of genomic DNA into DNA nanoballs. Unchained sequencing by ligation is then used to determine the nucleotide sequence. This method of DNA sequencing allows large numbers of DNA nanoballs to be sequenced per run.

Helicos's single-molecule sequencing uses DNA fragments with added polyA tail adapters, which are attached to the flow cell surface. The next steps involve extension-based sequencing with cyclic washes of the flow cell with fluorescently labeled nucleotides (one nucleotide type at a time, as with the Sanger method). The reads are performed by the Helioscope sequencer.

SMRT sequencing is based on the sequencing by synthesis approach. The DNA is synthesized in zero-mode wave-guides (ZMWs)—small well-like containers with the capturing tools located at the bottom of the well. The sequencing is performed with use of unmodified polymerase (attached to the ZMW bottom) and fluorescently labeled nucleotides flowing freely in the solution. The wells are constructed in a way that only the fluorescence occurring by the bottom of the well is detected. The fluorescent label is detached from the nucleotide at its incorporation into the DNA strand, leaving an unmodified DNA strand.

Single molecule real time sequencing based on RNA polymerase (RNAP), which is attached to a polystyrene bead, with distal end of sequenced DNA is attached to another bead, with both beads being placed in optical traps. RNAP motion during transcription brings the beads in closer and their relative distance changes, which can then be recorded at a single nucleotide resolution. The sequence is deduced based on the four readouts with lowered concentrations of each of the four nucleotide types (similarly to Sangers method).

Nanopore sequencing is based on the readout of electrical signal occurring at nucleotides passing by alpha-hemolysin pores covalently bound with cyclodextrin. The DNA passing through the nanopore changes its ion current. This change is dependent on the shape, size and length of the DNA sequence. Each type of the nucleotide blocks the ion flow through the pore for a different period of time.

VisiGen Biotechnologies uses a specially engineered DNA polymerase. This polymerase acts as a sensor—having incorporated a donor fluorescent dye by its active centre. This donor dye acts by FRET (fluorescent resonant energy transfer), inducing fluorescence of differently labeled nucleotides. This approach allows reads performed at the speed at which polymerase incorporates nucleotides into the sequence (several hundred per second). The nucleotide fluorochrome is released after the incorporation into the DNA strand.

Sequencing by hybridization is a non-enzymatic method that uses a DNA microarray. A single pool of DNA whose sequence is to be determined is fluorescently labeled and hybridized to an array containing known sequences. Strong hybridization signals from a given spot on the array identify its sequence in the DNA being sequenced. Mass spectrometry may be used to determine mass differences between DNA fragments produced in chain-termination reactions.

Another NGS approach is sequencing by synthesis (SBS) technology which is capable of overcoming the limitations of existing pyrosequencing based NGS platforms. Such technologies rely on complex enzymatic cascades for read out, are unreliable for the accurate determination of the number of nucleotides in homopolymeric regions and require excessive amounts of time to run individual nucleotides across growing DNA strands. The SBS NGS platform uses a direct sequencing approach to produce a sequencing strategy with very a high precision, rapid pace and low cost.

SBS sequencing is initialized by fragmenting of the template DNA into fragments, amplification, annealing of DNA sequencing primers, and finally affixing as a high-density array of spots onto a glass chip. The array of DNA fragments are sequenced by extending each fragment with modified nucleotides containing cleavable chemical moieties linked to fluorescent dyes capable of discriminating all four possible nucleotides. The array is scanned continuously by a high-resolution electronic camera (Measure) to determine the fluorescent intensity of each base (A, C, G or T) that was newly incorporated into the extended DNA fragment. After the incorporation of each modified base the array is exposed to cleavage chemistry to break off the fluorescent dye and end cap allowing additional bases to be added. The process is then repeated until the fragment is completely sequenced or maximal read length has been achieved.

mRNA Detection and Quantification

A number of widely used procedures exist for detecting and determining the abundance of a particular mRNA in a total or poly(A) RNA sample. For example, specific mRNAs can be detected using Northern blot analysis, nuclease protection assays (NPA), in situ hybridization (e.g., fluorescence in situ hybridization (FISH)), or reverse transcription-polymerase chain reaction (RT-PCR), and microarray.

In theory, each of these techniques can be used to detect specific RNAs and to precisely determine their expression level. In general, Northern analysis is the only method that provides information about transcript size, whereas NPAs are the easiest way to simultaneously examine multiple messages. In situ hybridization is used to localize expression of a particular gene within a tissue or cell type, and RT-PCR is the most sensitive method for detecting and quantitating gene expression.

RT-PCR allows for the detection of the RNA transcript of any gene, regardless of the scarcity of the starting material or relative abundance of the specific mRNA. In RT-PCR, an RNA template is copied into a complementary DNA (cDNA) using a retroviral reverse transcriptase. The cDNA is then amplified exponentially by PCR using a DNA polymerase. The reverse transcription and PCR reactions can occur in the same or difference tubes. RT-PCR is somewhat tolerant of degraded RNA. As long as the RNA is intact within the region spanned by the primers, the target will be amplified.

Relative quantitative RT-PCR involves amplifying an internal control simultaneously with the gene of interest. The internal control is used to normalize the samples. Once normalized, direct comparisons of relative abundance of a specific mRNA can be made across the samples. It is crucial to choose an internal control with a constant level of expression across all experimental samples (i.e., not affected by experimental treatment). Commonly used internal controls (e.g., GAPDH, β-actin, cyclophilin) often vary in expression and, therefore, may not be appropriate internal controls. Additionally, most common internal controls are expressed at much higher levels than the mRNA being studied. For relative RT-PCR results to be meaningful, all products of the PCR reaction must be analyzed in the linear range of amplification. This becomes difficult for transcripts of widely different levels of abundance.

Competitive RT-PCR is used for absolute quantitation. This technique involves designing, synthesizing, and accurately quantitating a competitor RNA that can be distinguished from the endogenous target by a small difference in size or sequence. Known amounts of the competitor RNA are added to experimental samples and RT-PCR is performed. Signals from the endogenous target are compared with signals from the competitor to determine the amount of target present in the sample.

Northern analysis is the easiest method for determining transcript size, and for identifying alternatively spliced transcripts and multigene family members. It can also be used to directly compare the relative abundance of a given message between all the samples on a blot. The Northern blotting procedure is straightforward and provides opportunities to evaluate progress at various points (e.g., intactness of the RNA sample and how efficiently it has transferred to the membrane). RNA samples are first separated by size via electrophoresis in an agarose gel under denaturing conditions. The RNA is then transferred to a membrane, crosslinked and hybridized with a labeled probe. Nonisotopic or high specific activity radiolabeled probes can be used including random-primed, nick-translated, or PCR-generated DNA probes, in vitro transcribed RNA probes, and oligonucleotides. Additionally, sequences with only partial homology (e.g., cDNA from a different species or genomic DNA fragments that might contain an exon) may be used as probes.

The Nuclease Protection Assay (NPA) (including both ribonuclease protection assays and Si nuclease assays) is a sensitive method for the detection and quantitation of specific mRNAs. The basis of the NPA is solution hybridization of an antisense probe (radiolabeled or nonisotopic) to an RNA sample. After hybridization, single-stranded, unhybridized probe and RNA are degraded by nucleases. The remaining protected fragments are separated on an acrylamide gel. Solution hybridization is typically more efficient than membrane-based hybridization, and it can accommodate up to 100 g of sample RNA, compared with the 20-30 μg maximum of blot hybridizations. NPAs are also less sensitive to RNA sample degradation than Northern analysis since cleavage is only detected in the region of overlap with the probe (probes are usually about 100-400 bases in length).

NPAs are the method of choice for the simultaneous detection of several RNA species. During solution hybridization and subsequent analysis, individual probe/target interactions are completely independent of one another. Thus, several RNA targets and appropriate controls can be assayed simultaneously (up to twelve have been used in the same reaction), provided that the individual probes are of different lengths. NPAs are also commonly used to precisely map mRNA termini and intron/exon junctions.

In situ hybridization (ISH) is a powerful and versatile tool for the localization of specific mRNAs in cells or tissues. Unlike Northern analysis and nuclease protection assays, ISH does not require the isolation or electrophoretic separation of RNA. Hybridization of the probe takes place within the cell or tissue. Since cellular structure is maintained throughout the procedure, ISH provides information about the location of mRNA within the tissue sample.

The procedure begins by fixing samples in neutral-buffered formalin, and embedding the tissue in paraffin. The samples are then sliced into thin sections and mounted onto microscope slides. (Alternatively, tissue can be sectioned frozen and post-fixed in paraformaldehyde.) After a series of washes to dewax and rehydrate the sections, a Proteinase K digestion is performed to increase probe accessibility, and a labeled probe is then hybridized to the sample sections. Radiolabeled probes are visualized with liquid film dried onto the slides, while non-isotopically labeled probes are conveniently detected with colorimetric or fluorescent reagents.

DNA Detection and Quantification

The methods, assays, and primer panels disclosed herein relate to the detection of nucleic acid variation that confer kinase inhibitor resistance in the form of, for example, point mutations and truncations of, KRAS, BRAF, KIT, EGFR, and/or ALK Thus, in one aspect, disclosed herein are methods, assays, and use of the disclosed primer panels for diagnosing an anaplastic lymphoma kinase (ALK) related cancer in a subject is resistant to a kinase inhibitor comprise performing NGS which sequences DNA from a tissue sample from the subject. It is understood that high throughput sequencing methods (also known as next generation sequencing methods) can comprise any known amplification and detection method for DNA known in the art.

A number of widely used procedures exist for detecting and determining the abundance of a particular DNA in a sample. For example, the technology of PCR permits amplification and subsequent detection of minute quantities of a target nucleic acid. Details of PCR are well described in the art, including, for example, U.S. Pat. No. 4,683,195 to Mullis et al., U.S. Pat. No. 4,683,202 to Mullis and U.S. Pat. No. 4,965,188 to Mullis et al. Generally, oligonucleotide primers are annealed to the denatured strands of a target nucleic acid, and primer extension products are formed by the polymerization of deoxynucleoside triphosphates by a polymerase. A typical PCR method involves repetitive cycles of template nucleic acid denaturation, primer annealing and extension of the annealed primers by the action of a thermostable polymerase. The process results in exponential amplification of the target nucleic acid, and thus allows the detection of targets existing in very low concentrations in a sample. It is understood and herein contemplated that there are variant PCR methods known in the art that may also be utilized in the disclosed methods, for example, Quantitative PCR (QPCR); microarrays, real-time PCR; hot start PCR; nested PCR; allele-specific PCR; and Touchdown PCR.

Microarrays

An array is an orderly arrangement of samples, providing a medium for matching known and unknown DNA samples based on base-pairing rules and automating the process of identifying the unknowns. An array experiment can make use of common assay systems such as microplates or standard blotting membranes, and can be created by hand or make use of robotics to deposit the sample. In general, arrays are described as macroarrays or microarrays, the difference being the size of the sample spots. Macroarrays contain sample spot sizes of about 300 microns or larger and can be easily imaged by existing gel and blot scanners. The sample spot sizes in microarray can be 300 microns or less, but typically less than 200 microns in diameter and these arrays usually contains thousands of spots. Microarrays require specialized robotics and/or imaging equipment that generally are not commercially available as a complete system. Terminologies that have been used in the literature to describe this technology include, but not limited to: biochip, DNA chip, DNA microarray, GENECHIP® (Affymetrix, Inc which refers to its high density, oligonucleotide-based DNA arrays), and gene array.

DNA microarrays, or DNA chips are fabricated by high-speed robotics, generally on glass or nylon substrates, for which probes with known identity are used to determine complementary binding, thus allowing massively parallel gene expression and gene discovery studies. An experiment with a single DNA chip can provide information on thousands of genes simultaneously. It is herein contemplated that the disclosed microarrays can be used to monitor gene expression, disease diagnosis, gene discovery, drug discovery (pharmacogenomics), and toxicological research or toxicogenomics.

There are two variants of the DNA microarray technology, in terms of the property of arrayed DNA sequence with known identity. Type I microarrays comprise a probe cDNA (500˜5,000 bases long) that is immobilized to a solid surface such as glass using robot spotting and exposed to a set of targets either separately or in a mixture. This method is traditionally referred to as DNA microarray. With Type I microarrays, localized multiple copies of one or more polynucleotide sequences, preferably copies of a single polynucleotide sequence are immobilized on a plurality of defined regions of the substrate's surface. A polynucleotide refers to a chain of nucleotides ranging from 5 to 10,000 nucleotides. These immobilized copies of a polynucleotide sequence are suitable for use as probes in hybridization experiments.

To prepare beads coated with immobilized probes, beads are immersed in a solution containing the desired probe sequence and then immobilized on the beads by covalent or non-covalent means. Alternatively, when the probes are immobilized on rods, a given probe can be spotted at defined regions of the rod. Typical dispensers include a micropipette delivering solution to the substrate with a robotic system to control the position of the micropipette with respect to the substrate. There can be a multiplicity of dispensers so that reagents can be delivered to the reaction regions simultaneously. In one embodiment, a microarray is formed by using ink-jet technology based on the piezoelectric effect, whereby a narrow tube containing a liquid of interest, such as oligonucleotide synthesis reagents, is encircled by an adapter. An electric charge sent across the adapter causes the adapter to expand at a different rate than the tube and forces a small drop of liquid onto a substrate.

Samples may be any sample containing polynucleotides (polynucleotide targets) of interest and obtained from any bodily fluid (blood, urine, saliva, phlegm, gastric juices, etc.), cultured cells, biopsies, or other tissue preparations. DNA or RNA can be isolated from the sample according to any of a number of methods well known to those of skill in the art. In one embodiment, total RNA is isolated using the TRIzol total RNA isolation reagent (Life Technologies, Inc., Rockville, Md.) and RNA is isolated using oligo d(T) column chromatography or glass beads. After hybridization and processing, the hybridization signals obtained should reflect accurately the amounts of control target polynucleotide added to the sample.

The plurality of defined regions on the substrate can be arranged in a variety of formats. For example, the regions may be arranged perpendicular or in parallel to the length of the casing. Furthermore, the targets do not have to be directly bound to the substrate, but rather can be bound to the substrate through a linker group. The linker groups may typically vary from about 6 to 50 atoms long. Linker groups include ethylene glycol oligomers, diamines, diacids and the like. Reactive groups on the substrate surface react with one of the terminal portions of the linker to bind the linker to the substrate. The other terminal portion of the linker is then functionalized for binding the probes.

Sample polynucleotides may be labeled with one or more labeling moieties to allow for detection of hybridized probe/target polynucleotide complexes. The labeling moieties can include compositions that can be detected by spectroscopic, photochemical, biochemical, bioelectronic, immunochemical, electrical, optical or chemical means. The labeling moieties include radioisotopes, such as ³²P, ³³P or ³⁵S, chemiluminescent compounds, labeled binding proteins, heavy metal atoms, spectroscopic markers, such as fluorescent markers and dyes, magnetic labels, linked enzymes, mass spectrometry tags, spin labels, electron transfer donors and acceptors, biotin, and the like.

Labeling can be carried out during an amplification reaction, such as polymerase chain reaction and in vitro or in vivo transcription reactions. Alternatively, the labeling moiety can be incorporated after hybridization once a probe-target complex his formed. In one embodiment, biotin is first incorporated during an amplification step as described above. After the hybridization reaction, unbound nucleic acids are rinsed away so that the only biotin remaining bound to the substrate is that attached to target polynucleotides that are hybridized to the polynucleotide probes. Then, an avidin-conjugated fluorophore, such as avidin-phycoerythrin, that binds with high affinity to biotin is added.

Hybridization causes a polynucleotide probe and a complementary target to form a stable duplex through base pairing. Hybridization methods are well known to those skilled in the art. Stringent conditions for hybridization can be defined by salt concentration, temperature, and other chemicals and conditions. Varying additional parameters, such as hybridization time, the concentration of detergent (sodium dodecyl sulfate, SDS) or solvent (formamide), and the inclusion or exclusion of carrier DNA, are well known to those skilled in the art. Additional variations on these conditions will be readily apparent to those skilled in the art.

Methods for detecting complex formation are well known to those skilled in the art. In one embodiment, the polynucleotide probes are labeled with a fluorescent label and measurement of levels and patterns of complex formation is accomplished by fluorescence microscopy, preferably confocal fluorescence microscopy. An argon ion laser excites the fluorescent label, emissions are directed to a photomultiplier and the amount of emitted light detected and quantitated. The detected signal should be proportional to the amount of probe/target polynucleotide complex at each position of the microarray. The fluorescence microscope can be associated with a computer-driven scanner device to generate a quantitative two-dimensional image of hybridization intensities. The scanned image is examined to determine the abundance/expression level of each hybridized target polynucleotide.

In a differential hybridization experiment, polynucleotide targets from two or more different biological samples are labeled with two or more different fluorescent labels with different emission wavelengths. Fluorescent signals are detected separately with different photomultipliers set to detect specific wavelengths. The relative abundances/expression levels of the target polynucleotides in two or more samples is obtained. Typically, microarray fluorescence intensities can be normalized to take into account variations in hybridization intensities when more than one microarray is used under similar test conditions. In one embodiment, individual polynucleotide probe/target complex hybridization intensities are normalized using the intensities derived from internal normalization controls contained on each microarray.

Type II microarrays comprise an array of oligonucleotides (20˜80-mer oligos) or peptide nucleic acid (PNA) probes that is synthesized either in situ (on-chip) or by conventional synthesis followed by on-chip immobilization. The array is exposed to labeled sample DNA, hybridized, and the identity/abundance of complementary sequences are determined. This method, “historically” called DNA chips, was developed at Affymetrix, Inc., which sells its photolithographically fabricated products under the GENECHIP® trademark.

The basic concept behind the use of Type II arrays for gene expression is simple: labeled cDNA or cRNA targets derived from the mRNA of an experimental sample are hybridized to nucleic acid probes attached to the solid support. By monitoring the amount of label associated with each DNA location, it is possible to infer the abundance of each mRNA species represented. Although hybridization has been used for decades to detect and quantify nucleic acids, the combination of the miniaturization of the technology and the large and growing amounts of sequence information, have enormously expanded the scale at which gene expression can be studied.

Microarray manufacturing can begin with a 5-inch square quartz wafer. Initially the quartz is washed to ensure uniform hydroxylation across its surface. Because quartz is naturally hydroxylated, it provides an excellent substrate for the attachment of chemicals, such as linker molecules, that are later used to position the probes on the arrays.

The wafer is placed in a bath of silane, which reacts with the hydroxyl groups of the quartz, and forms a matrix of covalently linked molecules. The distance between these silane molecules determines the probes' packing density, allowing arrays to hold over 500,000 probe locations, or features, within a mere 1.28 square centimeters. Each of these features harbors millions of identical DNA molecules. The silane film provides a uniform hydroxyl density to initiate probe assembly. Linker molecules, attached to the silane matrix, provide a surface that may be spatially activated by light.

Probe synthesis occurs in parallel, resulting in the addition of an A, C, T, or G nucleotide to multiple growing chains simultaneously. To define which oligonucleotide chains will receive a nucleotide in each step, photolithographic masks, carrying 18 to 20 square micron windows that correspond to the dimensions of individual features, are placed over the coated wafer. The windows are distributed over the mask based on the desired sequence of each probe. When ultraviolet light is shone over the mask in the first step of synthesis, the exposed linkers become deprotected and are available for nucleotide coupling.

Once the desired features have been activated, a solution containing a single type of deoxynucleotide with a removable protection group is flushed over the wafer's surface. The nucleotide attaches to the activated linkers, initiating the synthesis process.

Although each position in the sequence of an oligonucleotide can be occupied by 1 of 4 nucleotides, resulting in an apparent need for 25×4, or 100, different masks per wafer, the synthesis process can be designed to significantly reduce this requirement. Algorithms that help minimize mask usage calculate how to best coordinate probe growth by adjusting synthesis rates of individual probes and identifying situations when the same mask can be used multiple times.

Some of the key elements of selection and design are common to the production of all microarrays, regardless of their intended application. Strategies to optimize probe hybridization, for example, are invariably included in the process of probe selection. Hybridization under particular pH, salt, and temperature conditions can be optimized by taking into account melting temperatures and using empirical rules that correlate with desired hybridization behaviors.

To obtain a complete picture of a gene's activity, some probes are selected from regions shared by multiple splice or polyadenylation variants. In other cases, unique probes that distinguish between variants are favored. Inter-probe distance is also factored into the selection process.

A different set of strategies is used to select probes for genotyping arrays that rely on multiple probes to interrogate individual nucleotides in a sequence. The identity of a target base can be deduced using four identical probes that vary only in the target position, each containing one of the four possible bases.

Alternatively, the presence of a consensus sequence can be tested using one or two probes representing specific alleles. To genotype heterozygous or genetically mixed samples, arrays with many probes can be created to provide redundant information, resulting in unequivocal genotyping. In addition, generic probes can be used in some applications to maximize flexibility. Some probe arrays, for example, allow the separation and analysis of individual reaction products from complex mixtures, such as those used in some protocols to identify single nucleotide polymorphisms (SNPs).

Real-Time PCR

Real-time PCR monitors the fluorescence emitted during the reaction as an indicator of amplicon production during each PCR cycle (i.e., in real time) as opposed to the endpoint detection. The real-time progress of the reaction can be viewed in some systems. Real-time PCR does not detect the size of the amplicon and thus does not allow the differentiation between DNA and cDNA amplification, however, it is not influenced by non-specific amplification unless SYBR Green is used. Real-time PCR quantitation eliminates post-PCR processing of PCR products. This helps to increase throughput and reduce the chances of carryover contamination. Real-time PCR also offers a wide dynamic range of up to 10⁷-fold. Dynamic range of any assay determines how much target concentration can vary and still be quantified. A wide dynamic range means that a wide range of ratios of target and normaliser can be assayed with equal sensitivity and specificity. It follows that the broader the dynamic range, the more accurate the quantitation. When combined with RT-PCR, a real-time RT-PCR reaction reduces the time needed for measuring the amount of amplicon by providing for the visualization of the amplicon as the amplification process is progressing.

The real-time PCR system is based on the detection and quantitation of a fluorescent reporter. This signal increases in direct proportion to the amount of PCR product in a reaction. By recording the amount of fluorescence emission at each cycle, it is possible to monitor the PCR reaction during exponential phase where the first significant increase in the amount of PCR product correlates to the initial amount of target template. The higher the starting copy number of the nucleic acid target, the sooner a significant increase in fluorescence is observed. A significant increase in fluorescence above the baseline value measured during the 3-15 cycles can indicate the detection of accumulated PCR product.

A fixed fluorescence threshold is set significantly above the baseline that can be altered by the operator. The parameter C_(T) (threshold cycle) is defined as the cycle number at which the fluorescence emission exceeds the fixed threshold.

There are three main fluorescence-monitoring systems for DNA amplification: (1) hydrolysis probes; (2) hybridising probes; and (3) DNA-binding agents. Hydrolysis probes include TaqMan probes, molecular beacons and scorpions. They use the fluorogenic 5′ exonuclease activity of Taq polymerase to measure the amount of target sequences in cDNA samples.

TaqMan probes are oligonucleotides longer than the primers (20-30 bases long with a Tm value of 10° C. higher) that contain a fluorescent dye usually on the 5′ base, and a quenching dye (usually TAMRA) typically on the 3′ base. When irradiated, the excited fluorescent dye transfers energy to the nearby quenching dye molecule rather than fluorescing (this is called FRET=Förster or fluorescence resonance energy transfer). Thus, the close proximity of the reporter and quencher prevents emission of any fluorescence while the probe is intact. TaqMan probes are designed to anneal to an internal region of a PCR product. When the polymerase replicates a template on which a TaqMan probe is bound, its 5′ exonuclease activity cleaves the probe. This ends the activity of quencher (no FRET) and the reporter dye starts to emit fluorescence which increases in each cycle proportional to the rate of probe cleavage. Accumulation of PCR products is detected by monitoring the increase in fluorescence of the reporter dye (note that primers are not labelled). TaqMan assay uses universal thermal cycling parameters and PCR reaction conditions. Because the cleavage occurs only if the probe hybridises to the target, the origin of the detected fluorescence is specific amplification. The process ofhybridisation and cleavage does not interfere with the exponential accumulation of the product. One specific requirement for fluorogenic probes is that there be no G at the 5′ end. A ‘G’ adjacent to the reporter dye can quench reporter fluorescence even after cleavage.

Molecular beacons also contain fluorescent (FAM, TAMRA, TET, ROX) and quenching dyes (typically DABCYL) at either end but they are designed to adopt a hairpin structure while free in solution to bring the fluorescent dye and the quencher in close proximity for FRET to occur. They have two arms with complementary sequences that form a very stable hybrid or stem. The close proximity of the reporter and the quencher in this hairpin configuration suppresses reporter fluorescence. When the beacon hybridises to the target during the annealing step, the reporter dye is separated from the quencher and the reporter fluoresces (FRET does not occur). Molecular beacons remain intact during PCR and must rebind to target every cycle for fluorescence emission. This will correlate to the amount of PCR product available. All real-time PCR chemistries allow detection of multiple DNA species (multiplexing) by designing each probe/beacon with a spectrally unique fluor/quench pair as long as the platform is suitable for melting curve analysis if SYBR green is used. By multiplexing, the target(s) and endogenous control can be amplified in single tube.

With Scorpion probes, sequence-specific priming and PCR product detection is achieved using a single oligonucleotide. The Scorpion probe maintains a stem-loop configuration in the unhybridised state. The fluorophore is attached to the 5′ end and is quenched by a moiety coupled to the 3′ end. The 3′ portion of the stem also contains sequence that is complementary to the extension product of the primer. This sequence is linked to the 5′ end of a specific primer via a non-amplifiable monomer. After extension of the Scorpion primer, the specific probe sequence is able to bind to its complement within the extended amplicon thus opening up the hairpin loop. This prevents the fluorescence from being quenched and a signal is observed.

Another alternative is the double-stranded DNA binding dye chemistry, which quantitates the amplicon production (including non-specific amplification and primer-dimer complex) by the use of a non-sequence specific fluorescent intercalating agent (SYBR-green I or ethidium bromide). It does not bind to ssDNA. SYBR green is a fluorogenic minor groove binding dye that exhibits little fluorescence when in solution but emits a strong fluorescent signal upon binding to double-stranded DNA. Disadvantages of SYBR green-based real-time PCR include the requirement for extensive optimisation. Furthermore, non-specific amplifications require follow-up assays (melting point curve or dissociation analysis) for amplicon identification. The method has been used in HFE-C282Y genotyping. Another controllable problem is that longer amplicons create a stronger signal (if combined with other factors, this may cause CDC camera saturation, see below). Normally SYBR green is used in singleplex reactions, however when coupled with melting point analysis, it can be used for multiplex reactions.

The threshold cycle or the C_(T) value is the cycle at which a significant increase in ΔRn is first detected (for definition of ΔRn, see below). The threshold cycle is when the system begins to detect the increase in the signal associated with an exponential growth of PCR product during the log-linear phase. This phase provides the most useful information about the reaction (certainly more important than the end-point). The slope of the log-linear phase is a reflection of the amplification efficiency. The efficiency (Eff) of the reaction can be calculated by the formula: Eff=10^((−1/slope))−1. The efficiency of the PCR should be 90-100% (3.6>slope>3.1). A number of variables can affect the efficiency of the PCR. These factors include length of the amplicon, secondary structure and primer quality. Although valid data can be obtained that fall outside of the efficiency range, the qRT-PCR should be further optimised or alternative amplicons designed. For the slope to be an indicator of real amplification (rather than signal drift), there has to be an inflection point. This is the point on the growth curve when the log-linear phase begins. It also represents the greatest rate of change along the growth curve. (Signal drift is characterised by gradual increase or decrease in fluorescence without amplification of the product.) The important parameter for quantitation is the C_(T). The higher the initial amount of genomic DNA, the sooner accumulated product is detected in the PCR process, and the lower the C_(T) value. The threshold should be placed above any baseline activity and within the exponential increase phase (which looks linear in the log transformation). Some software allows determination of the cycle threshold (C_(T)) by a mathematical analysis of the growth curve. This provides better run-to-run reproducibility. A C_(T) value of 40 means no amplification and this value cannot be included in the calculations. Besides being used for quantitation, the C_(T) value can be used for qualitative analysis as a pass/fail measure.

Multiplex TaqMan assays can be performed using multiple dyes with distinct emission wavelengths. Available dyes for this purpose are FAM, TET, VIC and JOE (the most expensive). TAMRA is reserved as the quencher on the probe and ROX as the passive reference. For best results, the combination of FAM (target) and VIC (endogenous control) is recommended (they have the largest difference in emission maximum) whereas JOE and VIC should not be combined. It is important that if the dye layer has not been chosen correctly, the machine will still read the other dye's spectrum. For example, both VIC and FAM emit fluorescence in a similar range to each other and when doing a single dye, the wells should be labelled correctly. In the case of multiplexing, the spectral compensation for the post run analysis should be turned on (on ABI 7700: Instrument/Diagnostics/Advanced Options/Miscellaneous). Activating spectral compensation improves dye spectral resolution.

Nested PCR

The disclosed methods can further utilize nested PCR. Nested PCR increases the specificity of DNA amplification, by reducing background due to non-specific amplification of DNA. Two sets of primers are being used in two successive PCRs. In the first reaction, one pair of primers is used to generate DNA products, which besides the intended target, may still consist of non-specifically amplified DNA fragments. The product(s) are then used in a second PCR with a set of primers whose binding sites are completely or partially different from and located 3′ of each of the primers used in the first reaction. Nested PCR is often more successful in specifically amplifying long DNA fragments than conventional PCR, but it requires more detailed knowledge of the target sequences.

Primers and Probes

The disclosed methods and assays can use primers and probes. As used herein, “primers” are a subset of probes which are capable of supporting some type of enzymatic manipulation and which can hybridize with a target nucleic acid such that the enzymatic manipulation can occur. A primer can be made from any combination of nucleotides or nucleotide derivatives or analogs available in the art which do not interfere with the enzymatic manipulation.

As used herein, “probes” are molecules capable of interacting with a target nucleic acid, typically in a sequence specific manner, for example through hybridization. The hybridization of nucleic acids is well understood in the art and discussed herein. Typically a probe can be made from any combination of nucleotides or nucleotide derivatives or analogs available in the art.

Disclosed are assays and methods which include the use of primers and probes, as well as, the disclosed primer panels all of which are capable of interacting with the disclosed nucleic acids such as ALK (SEQ ID NO: 1), BRAF, EGFR, KIT, or KRAS or their complement. For example, any of the primers or primer sets from Table 7-14 can be used in the disclosed primer panels or any of the methods and assays disclosed herein. In certain embodiments the primers are used to support nucleic acid extension reactions, nucleic acid replication reactions, and/or nucleic acid amplification reactions. Typically the primers will be capable of being extended in a sequence specific manner. Extension of a primer in a sequence specific manner includes any methods wherein the sequence and/or composition of the nucleic acid molecule to which the primer is hybridized or otherwise associated directs or influences the composition or sequence of the product produced by the extension of the primer. Extension of the primer in a sequence specific manner therefore includes, but is not limited to, PCR, DNA sequencing, DNA extension, DNA polymerization, RNA transcription, or reverse transcription. Techniques and conditions that amplify the primer in a sequence specific manner are disclosed. In certain embodiments the primers are used for the DNA amplification reactions, such as PCR or direct sequencing. It is understood that in certain embodiments the primers can also be extended using non-enzymatic techniques, where for example, the nucleotides or oligonucleotides used to extend the primer are modified such that they will chemically react to extend the primer in a sequence specific manner. Typically the disclosed primers hybridize with the disclosed nucleic acids or region of the nucleic acids or they hybridize with the complement of the nucleic acids or complement of a region of the nucleic acids. As an example of the use of primers, one or more primers can be used to create extension products from and templated by a first nucleic acid.

The size of the primers or probes for interaction with the nucleic acids can be any size that supports the desired enzymatic manipulation of the primer, such as DNA amplification or the simple hybridization of the probe or primer. A typical primer or probe would be at least 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1250, 1500, 1750, 2000, 2250, 2500, 2750, 3000, 3500, or 4000 nucleotides long.

In other embodiments a primer or probe can be less than or equal to 6, 7, 8, 9, 10, 11, 12 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1250, 1500, 1750, 2000, 2250, 2500, 2750, 3000, 3500, or 4000 nucleotides long.

The primers for the nucleic acid of interest typically will be used to produce extension products and/or other replicated or amplified products that contain a region of the nucleic acid of interest. The size of the product can be such that the size can be accurately determined to within 3, or 2 or 1 nucleotides.

In certain embodiments the product can be, for example, at least 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1250, 1500, 1750, 2000, 2250, 2500, 2750, 3000, 3500, or 4000 nucleotides long.

In other embodiments the product can be, for example, less than or equal to 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1250, 1500, 1750, 2000, 2250, 2500, 2750, 3000, 3500, or 4000 nucleotides long.

It is understood and herein contemplated that there are situations where it may be advantageous to utilize more than one primer pair to detect the presence of mutations conferring inhibitor resistance in EGFR, BRAF, KIT, KRAS, or ALK. Such RT-PCR, real-time PCT or other PCR reactions can be conducted separately, or in a single reaction. When multiple primer pairs are placed into a single reaction, this is referred to as “multiplex PCR.” It is understood and herein contemplated that any combination of two or more or three or more the forward and/or reverse primers disclosed herein can be used in the multiplex reaction.

Fluorescent Change Probes and Primers

Fluorescent change probes and fluorescent change primers refer to all probes and primers that involve a change in fluorescence intensity or wavelength based on a change in the form or conformation of the probe or primer and nucleic acid to be detected, assayed or replicated. Examples of fluorescent change probes and primers include molecular beacons, Amplifluors, FRET probes, cleavable FRET probes, TaqMan probes, scorpion primers, fluorescent triplex oligos including but not limited to triplex molecular beacons or triplex FRET probes, fluorescent water-soluble conjugated polymers, PNA probes and QPNA probes.

Fluorescent change probes and primers can be classified according to their structure and/or function. Fluorescent change probes include hairpin quenched probes, cleavage quenched probes, cleavage activated probes, and fluorescent activated probes. Fluorescent change primers include stem quenched primers and hairpin quenched primers.

Hairpin quenched probes are probes that when not bound to a target sequence form a hairpin structure (and, typically, a loop) that brings a fluorescent label and a quenching moiety into proximity such that fluorescence from the label is quenched. When the probe binds to a target sequence, the stem is disrupted, the quenching moiety is no longer in proximity to the fluorescent label and fluorescence increases. Examples of hairpin quenched probes are molecular beacons, fluorescent triplex oligos, triplex molecular beacons, triplex FRET probes, and QPNA probes.

Cleavage activated probes are probes where fluorescence is increased by cleavage of the probe. Cleavage activated probes can include a fluorescent label and a quenching moiety in proximity such that fluorescence from the label is quenched. When the probe is clipped or digested (typically by the 5′-3′ exonuclease activity of a polymerase during amplification), the quenching moiety is no longer in proximity to the fluorescent label and fluorescence increases. TaqMan probes are an example of cleavage activated probes.

Cleavage quenched probes are probes where fluorescence is decreased or altered by cleavage of the probe. Cleavage quenched probes can include an acceptor fluorescent label and a donor moiety such that, when the acceptor and donor are in proximity, fluorescence resonance energy transfer from the donor to the acceptor causes the acceptor to fluoresce. The probes are thus fluorescent, for example, when hybridized to a target sequence. When the probe is clipped or digested (typically by the 5′-3′ exonuclease activity of a polymerase during amplification), the donor moiety is no longer in proximity to the acceptor fluorescent label and fluorescence from the acceptor decreases. If the donor moiety is itself a fluorescent label, it can release energy as fluorescence (typically at a different wavelength than the fluorescence of the acceptor) when not in proximity to an acceptor. The overall effect would then be a reduction of acceptor fluorescence and an increase in donor fluorescence. Donor fluorescence in the case of cleavage quenched probes is equivalent to fluorescence generated by cleavage activated probes with the acceptor being the quenching moiety and the donor being the fluorescent label. Cleavable FRET (fluorescence resonance energy transfer) probes are an example of cleavage quenched probes.

Fluorescent activated probes are probes or pairs of probes where fluorescence is increased or altered by hybridization of the probe to a target sequence. Fluorescent activated probes can include an acceptor fluorescent label and a donor moiety such that, when the acceptor and donor are in proximity (when the probes are hybridized to a target sequence), fluorescence resonance energy transfer from the donor to the acceptor causes the acceptor to fluoresce. Fluorescent activated probes are typically pairs of probes designed to hybridize to adjacent sequences such that the acceptor and donor are brought into proximity. Fluorescent activated probes can also be single probes containing both a donor and acceptor where, when the probe is not hybridized to a target sequence, the donor and acceptor are not in proximity but where the donor and acceptor are brought into proximity when the probe hybridized to a target sequence. This can be accomplished, for example, by placing the donor and acceptor on opposite ends of the probe and placing target complement sequences at each end of the probe where the target complement sequences are complementary to adjacent sequences in a target sequence. If the donor moiety of a fluorescent activated probe is itself a fluorescent label, it can release energy as fluorescence (typically at a different wavelength than the fluorescence of the acceptor) when not in proximity to an acceptor (that is, when the probes are not hybridized to the target sequence). When the probes hybridize to a target sequence, the overall effect would then be a reduction of donor fluorescence and an increase in acceptor fluorescence. FRET probes are an example of fluorescent activated probes.

Stem quenched primers are primers that when not hybridized to a complementary sequence form a stem structure (either an intramolecular stem structure or an intermolecular stem structure) that brings a fluorescent label and a quenching moiety into proximity such that fluorescence from the label is quenched. When the primer binds to a complementary sequence, the stem is disrupted; the quenching moiety is no longer in proximity to the fluorescent label and fluorescence increases. In the disclosed method, stem quenched primers are used as primers for nucleic acid synthesis and thus become incorporated into the synthesized or amplified nucleic acid. Examples of stem quenched primers are peptide nucleic acid quenched primers and hairpin quenched primers.

Peptide nucleic acid quenched primers are primers associated with a peptide nucleic acid quencher or a peptide nucleic acid fluor to form a stem structure. The primer contains a fluorescent label or a quenching moiety and is associated with either a peptide nucleic acid quencher or a peptide nucleic acid fluor, respectively. This puts the fluorescent label in proximity to the quenching moiety. When the primer is replicated, the peptide nucleic acid is displaced, thus allowing the fluorescent label to produce a fluorescent signal.

Hairpin quenched primers are primers that when not hybridized to a complementary sequence form a hairpin structure (and, typically, a loop) that brings a fluorescent label and a quenching moiety into proximity such that fluorescence from the label is quenched. When the primer binds to a complementary sequence, the stem is disrupted; the quenching moiety is no longer in proximity to the fluorescent label and fluorescence increases. Hairpin quenched primers are typically used as primers for nucleic acid synthesis and thus become incorporated into the synthesized or amplified nucleic acid. Examples of hairpin quenched primers are Amplifluor primers and scorpion primers.

Cleavage activated primers are similar to cleavage activated probes except that they are primers that are incorporated into replicated strands and are then subsequently cleaved.

Labels

To aid in detection and quantitation of nucleic acids produced using the disclosed methods, labels can be directly incorporated into nucleotides and nucleic acids or can be coupled to detection molecules such as probes and primers. As used herein, a label is any molecule that can be associated with a nucleotide or nucleic acid, directly or indirectly, and which results in a measurable, detectable signal, either directly or indirectly. Many such labels for incorporation into nucleotides and nucleic acids or coupling to nucleic acid probes are known to those of skill in the art. Examples of labels suitable for use in the disclosed method are radioactive isotopes, fluorescent molecules, phosphorescent molecules, enzymes, antibodies, and ligands. Fluorescent labels, especially in the context of fluorescent change probes and primers, are useful for real-time detection of amplification.

Examples of suitable fluorescent labels include fluorescein isothiocyanate (FITC), 5,6-carboxymethyl fluorescein, Texas red, nitrobenz-2-oxa-1,3-diazol-4-yl (NBD), coumarin, dansyl chloride, rhodamine, amino-methyl coumarin (AMCA), Eosin, Erythrosin, BODIPY®, CASCADE BLUE®, OREGON GREEN®, pyrene, lissamine, xanthenes, acridines, oxazines, phycoerythrin, macrocyclic chelates of lanthanide ions such as quantum Dye™, fluorescent energy transfer dyes, such as thiazole orange-ethidium heterodimer, and the cyanine dyes Cy3, Cy3.5, Cy5, Cy5.5 and Cy7. Examples of other specific fluorescent labels include 3-Hydroxypyrene 5,8,10-Tri Sulfonic acid, 5-Hydroxy Tryptamine (5-HT), Acid Fuchsin, Alizarin Complexon, Alizarin Red, Allophycocyanin, Aminocoumarin, Anthroyl Stearate, Astrazon Brilliant Red 4G, Astrazon Orange R, Astrazon Red 6B, Astrazon Yellow 7 GLL, Atabrine, Auramine, Aurophosphine, Aurophosphine G, BAO 9 (Bisaminophenyloxadiazole), BCECF, BerberineSulphate, Bisbenzamide, Blancophor FFG Solution, Blancophor SV, Bodipy F1, Brilliant Sulphoflavin FF, Calcien Blue, Calcium Green, Calcofluor RW Solution, Calcofluor White, Calcophor White ABT Solution, Calcophor White Standard Solution, Carbostyryl, Cascade Yellow, Catecholamine, Chinacrine, Coriphosphine O, Coumarin-Phalloidin, CY3.1 8, CY5.1 8, CY7, Dans (1-Dimethyl Amino Naphaline 5 Sulphonic Acid), Dansa (DiaminoNaphtylSulphonic Acid), Dansyl NH-CH3, Diamino Phenyl Oxydiazole (DAO), Dimethylamino-5-Sulphonic acid, DipyrrometheneboronDifluoride, Diphenyl Brilliant Flavine 7GFF, Dopamine, Erythrosin ITC, Euchrysin, FIF (Formaldehyde Induced Fluorescence), Flazo Orange, Fluo 3, Fluorescamine, Fura-2, Genacryl Brilliant Red B, Genacryl Brilliant Yellow 10GF, Genacryl Pink 3G, Genacryl Yellow 5GF, Gloxalic Acid, Granular Blue, Haematoporphyrin, Indo-1, IntrawhiteCf Liquid, Leucophor PAF, Leucophor SF, Leucophor WS, LissamineRhodamine B200 (RD200), Lucifer Yellow CH, Lucifer Yellow VS, Magdala Red, Marina Blue, Maxilon Brilliant Flavin 10 GFF, Maxilon Brilliant Flavin 8 GFF, MPS (Methyl Green PyronineStilbene), Mithramycin, NBD Amine, Nitrobenzoxadidole, Noradrenaline, Nuclear Fast Red, Nuclear Yellow, Nylosan Brilliant Flavin E8G, Oxadiazole, Pacific Blue, Pararosaniline (Feulgen), Phorwite AR Solution, Phorwite BKL, Phorwite Rev, Phorwite RPA, Phosphine 3R, Phthalocyanine, Phycoerythrin R, Phycoerythrin B, PolyazaindacenePontochrome Blue Black, Porphyrin, Primuline, Procion Yellow, Pyronine, Pyronine B, Pyrozal Brilliant Flavin 7GF, Quinacrine Mustard, Rhodamine 123, Rhodamine 5 GLD, Rhodamine 6G, Rhodamine B, Rhodamine B 200, Rhodamine B Extra, Rhodamine BB, Rhodamine BG, Rhodamine WT, Serotonin, Sevron Brilliant Red 2B, Sevron Brilliant Red 4G, Sevron Brilliant Red B, Sevron Orange, Sevron Yellow L, SITS (Primuline), SITS (Stilbenelsothiosulphonic acid), Stilbene, Snarf 1, sulphoRhodamine B Can C, SulphoRhodamine G Extra, Tetracycline, Thiazine Red R, Thioflavin S, Thioflavin TCN, Thioflavin 5, Thiolyte, Thiozol Orange, Tinopol CBS, True Blue, Ultralite, Uranine B, Uvitex SFC, Xylene Orange, and XRITC.

The absorption and emission maxima, respectively, for some of these fluors are: FITC (490 nm; 520 nm), Cy3 (554 nm; 568 nm), Cy3.5 (581 nm; 588 nm), Cy5 (652 nm: 672 nm), Cy5.5 (682 nm; 703 nm) and Cy7 (755 nm; 778 nm), thus allowing their simultaneous detection. Other examples of fluorescein dyes include 6-carboxyfluorescein (6-FAM), 2′,4′,1,4,-tetrachlorofluorescein (TET), 2′,4′,5′,7′,1,4-hexachlorofluorescein (HEX), 2′,7′-dimethoxy-4′,5′-dichloro-6-carboxyrhodamine (JOE), 2′-chloro-5′-fluoro-7′,8′-fused phenyl-1,4-dichloro-6-carboxyfluorescein (NED), and 2′-chloro-7′-phenyl-1,4-dichloro-6-carboxyfluorescein (VIC). Fluorescent labels can be obtained from a variety of commercial sources, including Amersham Pharmacia Biotech, Piscataway, N.J.; Molecular Probes, Eugene, Oreg.; and Research Organics, Cleveland, Ohio.

Additional labels of interest include those that provide for signal only when the probe with which they are associated is specifically bound to a target molecule, where such labels include: “molecular beacons” as described in Tyagi & Kramer, Nature Biotechnology (1996) 14:303 and EP 0 070 685 Bi. Other labels of interest include those described in U.S. Pat. No. 5,563,037 which is incorporated herein by reference.

Labeled nucleotides are a form of label that can be directly incorporated into the amplification products during synthesis. Examples of labels that can be incorporated into amplified nucleic acids include nucleotide analogs such as BrdUrd, aminoallyldeoxyuridine, 5-methylcytosine, bromouridine, and nucleotides modified with biotin or with suitable haptens such as digoxygenin. Suitable fluorescence-labeled nucleotides are Fluorescein-isothiocyanate-dUTP, Cyanine-3-dUTP and Cyanine-5-dUTP. One example of a nucleotide analog label for DNA is BrdUrd (bromodeoxyuridine, BrdUrd, BrdU, BUdR, Sigma-Aldrich Co). Other examples of nucleotide analogs for incorporation of label into DNA are AA-dUTP (aminoallyl-deoxyuridine triphosphate, Sigma-Aldrich Co.), and 5-methyl-dCTP (Roche Molecular Biochemicals). One example of a nucleotide analog for incorporation of label into RNA is biotin-16-UTP (biotin-16-uridine-5′-triphosphate, Roche Molecular Biochemicals). Fluorescein, Cy3, and Cy5 can be linked to dUTP for direct labeling. Cy3.5 and Cy7 are available as avidin or anti-digoxygenin conjugates for secondary detection of biotin- or digoxygenin-labeled probes.

Labels that are incorporated into amplified nucleic acid, such as biotin, can be subsequently detected using sensitive methods well-known in the art. For example, biotin can be detected using streptavidin-alkaline phosphatase conjugate (Tropix, Inc.), which is bound to the biotin and subsequently detected by chemiluminescence of suitable substrates (for example, chemiluminescent substrate CSPD: disodium, 3-(4-methoxyspiro-[1,2,-dioxetane-3-2′-(5′-chloro)tricyclo[3.3.1.1³′7]decane]-4-yl) phenyl phosphate; Tropix, Inc.). Labels can also be enzymes, such as alkaline phosphatase, soybean peroxidase, horseradish peroxidase and polymerases, that can be detected, for example, with chemical signal amplification or by using a substrate to the enzyme which produces light (for example, a chemiluminescent 1,2-dioxetane substrate) or fluorescent signal.

Molecules that combine two or more of these labels are also considered labels. Any of the known labels can be used with the disclosed probes, tags, and method to label and detect nucleic acid amplified using the disclosed method. Methods for detecting and measuring signals generated by labels are also known to those of skill in the art. For example, radioactive isotopes can be detected by scintillation counting or direct visualization; fluorescent molecules can be detected with fluorescent spectrophotometers; phosphorescent molecules can be detected with a spectrophotometer or directly visualized with a camera; enzymes can be detected by detection or visualization of the product of a reaction catalyzed by the enzyme; antibodies can be detected by detecting a secondary label coupled to the antibody. As used herein, detection molecules are molecules which interact with amplified nucleic acid and to which one or more labels are coupled.

The disclosed methods, assays, and primer panels can be used to diagnose any disease where uncontrolled cellular proliferation occurs herein referred to as “cancer”. A non-limiting list of different types of ALK related cancers is as follows: lymphomas (Hodgkins and non-Hodgkins), leukemias, carcinomas, carcinomas of solid tissues, squamous cell carcinomas, adenocarcinomas, sarcomas, gliomas, high grade gliomas, blastomas, neuroblastomas, plasmacytomas, histiocytomas, melanomas, adenomas, hypoxic tumours, myelomas, AIDS-related lymphomas or sarcomas, metastatic cancers, or cancers in general. In particular, the disclosed methods, assays, and kits relate to the diagnosis, detection, or prognosis of inflammatory breast cancer

A representative but non-limiting list of cancers that the disclosed methods can be used to diagnose is the following: lymphoma, B cell lymphoma (including diffuse large B-cell lymphoma), B-cell plasmablastic/immunoblastic lymphomas, T cell lymphoma (including T- or null-cell lymphomas), mycosis fungoides, Hodgkin's Disease, myeloid leukemia, bladder cancer, brain cancer, nervous system cancer, head and neck cancer, squamous cell carcinoma of head and neck, kidney cancer, lung cancers such as small cell lung cancer and non-small cell lung cancer, neuroblastoma/glioblastoma, anaplastic large-cell lymphoma (ALCL), inflammatory myofibroblastic tumors, ovarian cancer, pancreatic cancer, prostate cancer, skin cancer, liver cancer, melanoma, malignant histiocytosis, squamous cell carcinomas of the mouth, throat, larynx, and lung, colon cancer, cervical cancer, cervical carcinoma, breast cancer (including inflammatory breast cancer), and epithelial cancer, renal cancer, genitourinary cancer, pulmonary cancer, esophageal squamous cell carcinoma, head and neck carcinoma, large bowel cancer, hematopoietic cancers; testicular cancer; colon and rectal cancers, prostatic cancer, or pancreatic cancer.

Nucleic Acids

The disclosed method and compositions make use of various nucleic acids. Generally, any nucleic acid can be used in the disclosed method. For example, the disclosed nucleic acids of interest and the disclosed reference nucleic acids can be chosen based on the desired analysis and information that is to be obtained or assessed. The disclosed methods also produce new and altered nucleic acids. The nature and structure of such nucleic acids will be established by the manner in which they are produced and manipulated in the methods. Thus, for example, extension products and hybridizing nucleic acids are produced in the disclosed methods. As used herein, hybridizing nucleic acids are hybrids of extension products and the second nucleic acid.

It is understood and contemplated herein that a nucleic acid of interest can be any nucleic acid to which the determination of the presence or absence of nucleotide variation is desired. Thus, for example, the nucleic acid of interest can comprise a sequence that corresponds to the wild-type sequence of the reference nucleic acid. It is further disclosed herein that the disclosed methods can be performed where the first nucleic acid is a reference nucleic acid and the second nucleic acid is a nucleic acid of interest or where the first nucleic acid is the nucleic acid of interest and the second nucleic acid is the reference nucleic acid.

It is understood and herein contemplated that a reference nucleic acid can be any nucleic acid against which a nucleic acid of interest is to be compared. Typically, the reference nucleic acid has a known sequence (and/or is known to have a sequence of interest as a reference). Although not required, it is useful if the reference sequence has a known or suspected close relationship to the nucleic acid of interest. For example, if a single nucleotide variation is desired to be detected, the reference sequence can be usefully chosen to be a sequence that is a homolog or close match to the nucleic acid of interest, such as a nucleic acid derived from the same gene or genetic element from the same or a related organism or individual. Thus, for example, it is contemplated herein that the reference nucleic acid can comprise a wild-type sequence or alternatively can comprise a known mutation including, for example, a mutation the presence or absence of which is associated with a disease or resistance to therapeutic treatment. Thus, for example, it is contemplated that the disclosed methods can be used to detect or diagnose the presence of known mutations in a nucleic acid of interest by comparing the nucleic acid of interest to a reference nucleic acid that comprises a wild-type sequence (i.e., is known not to possess the mutation) and examining for the presence or absence of variation in the nucleic acid of interest, where the absence of variation would indicate the absence of a mutation. Alternatively, the reference nucleic acid can possess a known mutation. Thus, for example, it is contemplated that the disclosed methods can be used to detect susceptibility for a disease or condition by comparing the nucleic acid of interest to a reference nucleic acid comprising a known mutation that indicates susceptibility for a disease and examining for the presence or absence of the mutation, wherein the presence of the mutation indicates a disease.

Herein, the term “nucleotide variation” refers to any change or difference in the nucleotide sequence of a nucleic acid of interest relative to the nucleotide sequence of a reference nucleic acid. Thus, a nucleotide variation is said to occur when the sequences between the reference nucleic acid and the nucleic acid of interest (or its complement, as appropriate in context) differ. Thus, for example, a substitution of an adenine (A) to a guanine (G) at a particular position in a nucleic acid would be a nucleotide variation provided the reference nucleic acid comprised an A at the corresponding position. It is understood and herein contemplated that the determination of a variation is based upon the reference nucleic acid and does not indicate whether or not a sequence is wild-type. Thus, for example, when a nucleic acid with a known mutation is used as the reference nucleic acid, a nucleic acid not possessing the mutation (including a wild-type nucleic acid) would be considered to possess a nucleotide variation (relative to the reference nucleic acid).

Nucleotides

The disclosed methods and compositions make use of various nucleotides. Throughout this application and the methods disclosed herein reference is made to the type of base for a nucleotide. It is understood and contemplated herein that where reference is made to a type of base, this refers a base that in a nucleotide in a nucleic acid strand is capable of hybridizing (binding) to a defined set of one or more of the canonical bases. Thus, for example, where reference is made to extension products extended in the presence of three types of nuclease resistant nucleotides and not in the presence of nucleotides that comprise the same type of base as the modified nucleotides, this means that if, for example, the base of the modified nucleotide was an adenine (A), the nuclease-resistant nucleotides can be, for example, guanine (G), thymine (T), and cytosine (C). Each of these bases (which represent the four canonical bases) is capable of hybridizing to a different one of the four canonical bases and thus each qualify as a different type of base as defined herein. As another example, inosine base pairs primarily with adenine and cytosine (in DNA) and thus can be considered a different type of base from adenine and from cytosine—which base pair with thymine and guanine, respectively—but not a different type of base from guanine or thymine-which base pair with cytosine and adenine, respectively-because the base pairing of guanine and thymine overlaps (that is, is not different from) the hybridization pattern of inosine.

Any type of modified or alternative base can be used in the disclosed methods and compositions, generally limited only by the capabilities of the enzymes used to use such bases. Many modified and alternative nucleotides and bases are known, some of which are described below and elsewhere herein. The type of base that such modified and alternative bases represent can be determined by the pattern of base pairing for that base as described herein. Thus for example, if the modified nucleotide was adenine, any analog, derivative, modified, or variant base that based pairs primarily with thymine would be considered the same type of base as adenine. In other words, so long as the analog, derivative, modified, or variant has the same pattern of base pairing as another base, it can be considered the same type of base. Modifications can made to the sugar or phosphate groups of a nucleotide. Generally such modifications will not change the base pairing pattern of the base. However, the base pairing pattern of a nucleotide in a nucleic acid strand determines the type of base of the base in the nucleotide.

Modified nucleotides to be incorporated into extension products and to be selectively removed by the disclosed agents in the disclosed methods can be any modified nucleotide that functions as needed in the disclosed method as is described elsewhere herein. Modified nucleotides can also be produced in existing nucleic acid strands, such as extension products, by, for example, chemical modification, enzymatic modification, or a combination.

Many types of nuclease-resistant nucleotides are known and can be used in the disclosed methods. For example, nucleotides have modified phosphate groups and/or modified sugar groups can be resistant to one or more nucleases. Nuclease-resistance is defined herein as resistance to removal from a nucleic acid by any one or more nucleases. Generally, nuclease resistance of a particular nucleotide can be defined in terms of a relevant nuclease. Thus, for example, if a particular nuclease is used in the disclosed method, the nuclease-resistant nucleotides need only be resistant to that particular nuclease. Examples of useful nuclease-resistant nucleotides include thio-modified nucleotides and borano-modified nucleotides.

There are a variety of molecules disclosed herein that are nucleic acid based. Non-limiting examples of these and other molecules are discussed herein. It is understood that for example, a nucleotide is a molecule that contains a base moiety, a sugar moiety and a phosphate moiety. Nucleotides can be linked together through their phosphate moieties and sugar moieties creating an intemucleoside linkage. The base moiety of a nucleotide can be adenine-9-yl (adenine, A), cytosine-1-yl (cytosine, C), guanine-9-yl (guanine, G), uracil-1-yl (uracil, U), and thymin-1-yl (thymine, T). The sugar moiety of a nucleotide is a ribose or a deoxyribose. The phosphate moiety of a nucleotide is pentavalent phosphate. A non-limiting example of a nucleotide would be 3′-AMP (3′-adenosine monophosphate) or 5′-GMP (5′-guanosine monophosphate).

A nucleotide analog is a nucleotide which contains some type of modification to either the base, sugar, or phosphate moieties. Modifications to the base moiety would include natural and synthetic modifications of A, C, G, and T/U as well as different purine or pyrimidine bases, such as uracil-5-yl (w), hypoxanthin-9-yl (inosine, I), and 2-aminoadenin-9-yl. A modified base includes but is not limited to 5-methylcytosine (5-me-C), 5-hydroxymethyl cytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-methyl and other alkyl derivatives of adenine and guanine, 2-propyl and other alkyl derivatives of adenine and guanine, 2-thiouracil, 2-thiothymine and 2-thiocytosine, 5-halouracil and cytosine, 5-propynyl uracil and cytosine, 6-azo uracil, cytosine and thymine, 5-uracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl and other 8-substituted adenines and guanines, 5-halo particularly 5-bromo, 5-trifluoromethyl and other 5-substituted uracils and cytosines, 7-methylguanine and 7-methyladenine, 8-azaguanine and 8-azaadenine, 7-deazaguanine and 7-deazaadenine and 3-deazaguanine and 3-deazaadenine. Additional base modifications can be found for example in U.S. Pat. No. 3,687,808, which is incorporated herein in its entirety for its teachings of base modifications. Certain nucleotide analogs, such as 5-substituted pyrimidines, 6-azapyrimidines and N-2, N-6 and O-6 substituted purines, including 2-aminopropyladenine, 5-propynyluracil and 5-propynylcytosine. 5-methylcytosine can increase the stability of duplex formation. Often time base modifications can be combined with for example a sugar modification, such as 2′-O-methoxyethyl, to achieve unique properties such as increased duplex stability.

Nucleotide analogs can also include modifications of the sugar moiety. Modifications to the sugar moiety would include natural modifications of the ribose and deoxy ribose as well as synthetic modifications. Sugar modifications include but are not limited to the following modifications at the 2′ position: OH; F; O-, S-, or N-alkyl; O-, S-, or N-alkenyl; O-, S- or N-alkynyl; or O-alkyl-O-alkyl, wherein the alkyl, alkenyl and alkynyl may be substituted or unsubstituted C1 to C10, alkyl or C2 to C10 alkenyl and alkynyl. 2′ sugar modifications also include but are not limited to —O[(CH2)n O]m CH3, —O(CH2)n OCH3, —O(CH2)n NH2, —O(CH2)n CH3, —O(CH2)n—ONH2, and —O(CH2)nON[(CH2)n CH3)]2, where n and m are from 1 to about 10.

Other modifications at the 2′ position include but are not limited to: C1 to C10 lower alkyl, substituted lower alkyl, alkaryl, aralkyl, O-alkaryl or O-aralkyl, SH, SCH3, OCN, Cl, Br, CN, CF3, OCF3, SOCH3, SO2 CH3, ONO2, NO2, N3, NH2, heterocycloalkyl, heterocycloalkaryl, aminoalkylamino, polyalkylamino, substituted silyl, an RNA cleaving group, a reporter group, an intercalator, a group for improving the pharmacokinetic properties of an oligonucleotide, or a group for improving the pharmacodynamic properties of an oligonucleotide, and other substituents having similar properties. Similar modifications may also be made at other positions on the sugar, particularly the 3′ position of the sugar on the 3′ terminal nucleotide or in 2′-5′ linked oligonucleotides and the 5′ position of 5′ terminal nucleotide. Modified sugars would also include those that contain modifications at the bridging ring oxygen, such as CH2 and S. Nucleotide sugar analogs may also have sugar mimetics such as cyclobutyl moieties in place of the pentofuranosyl sugar.

Nucleotide analogs can also be modified at the phosphate moiety. Modified phosphate moieties include but are not limited to those that can be modified so that the linkage between two nucleotides contains a phosphorothioate, chiral phosphorothioate, phosphorodithioate, phosphotriester, aminoalkylphosphotriester, methyl and other alkyl phosphonates including 3′-alkylene phosphonate and chiral phosphonates, phosphinates, phosphoramidates including 3′-amino phosphoramidate and aminoalkylphosphoramidates, thionophosphoramidates, thionoalkylphosphonates, thionoalkylphosphotriesters, and boranophosphates. It is understood that these phosphate or modified phosphate linkage between two nucleotides can be through a 3′-5′ linkage or a 2′-5′ linkage, and the linkage can contain inverted polarity such as 3′-5′ to 5′-3′ or 2′-5′ to 5′-2′. Various salts, mixed salts and free acid forms are also included.

It is understood that nucleotide analogs need only contain a single modification, but may also contain multiple modifications within one of the moieties or between different moieties.

Nucleotide substitutes are molecules having similar functional properties to nucleotides, but which do not contain a phosphate moiety, such as peptide nucleic acid (PNA). Nucleotide substitutes are molecules that will recognize nucleic acids in a Watson-Crick or Hoogsteen manner, but which are linked together through a moiety other than a phosphate moiety. Nucleotide substitutes are able to conform to a double helix type structure when interacting with the appropriate target nucleic acid.

Nucleotide substitutes are nucleotides or nucleotide analogs that have had the phosphate moiety and/or sugar moieties replaced. Nucleotide substitutes do not contain a standard phosphorus atom. Substitutes for the phosphate can be for example, short chain alkyl or cycloalkylinternucleoside linkages, mixed heteroatom and alkyl or cycloalkylinternucleoside linkages, or one or more short chain heteroatomic or heterocyclic internucleoside linkages. These include those having morpholino linkages (formed in part from the sugar portion of a nucleoside); siloxane backbones; sulfide, sulfoxide and sulfone backbones; formacetyl and thioformacetyl backbones; methylene formacetyl and thioformacetyl backbones; alkene containing backbones; sulfamate backbones; methyleneimino and methylenehydrazino backbones; sulfonate and sulfonamide backbones; amide backbones; and others having mixed N, O, S and CH2 component parts.

It is also understood in a nucleotide substitute that both the sugar and the phosphate moieties of the nucleotide can be replaced, by for example an amide type linkage (aminoethylglycine) (PNA).

It is also possible to link other types of molecules (conjugates) to nucleotides or nucleotide analogs. Conjugates can be chemically linked to the nucleotide or nucleotide analogs. Such conjugates include but are not limited to lipid moieties such as a cholesterol moiety, cholic acid, a thioether, e.g., hexyl-S-tritylthiol, a thiocholesterol, an aliphatic chain, e.g., dodecandiol or undecyl residues, a phospholipid, e.g., di-hexadecyl-rac-glycerol or triethylammonium 1,2-di-O-hexadecyl-rac-glycero-3-H-phosphonate, a polyamine or a polyethylene glycol chain, or adamantane acetic acid, a palmityl moiety, or an octadecylamine or hexylamino-carbonyl-oxycholesterol moiety.

A Watson-Crick interaction is at least one interaction with the Watson-Crick face of a nucleotide, nucleotide analog, or nucleotide substitute. The Watson-Crick face of a nucleotide, nucleotide analog, or nucleotide substitute includes the C2, Ni, and C6 positions of a purine based nucleotide, nucleotide analog, or nucleotide substitute and the C2, N3, C4 positions of a pyrimidine based nucleotide, nucleotide analog, or nucleotide substitute.

A Hoogsteen interaction is the interaction that takes place on the Hoogsteen face of a nucleotide or nucleotide analog, which is exposed in the major groove of duplex DNA. The Hoogsteen face includes the N7 position and reactive groups (NH2 or O) at the C6 position of purine nucleotides.

Hybridization/Selective Hybridization

The term hybridization typically means a sequence driven interaction between at least two nucleic acid molecules, such as a primer or a probe and a gene. Sequence driven interaction means an interaction that occurs between two nucleotides or nucleotide analogs or nucleotide derivatives in a nucleotide specific manner. For example, G interacting with C or A interacting with T are sequence driven interactions. Typically sequence driven interactions occur on the Watson-Crick face or Hoogsteen face of the nucleotide. The hybridization of two nucleic acids is affected by a number of conditions and parameters known to those of skill in the art. For example, the salt concentrations, pH, and temperature of the reaction all affect whether two nucleic acid molecules will hybridize.

Parameters for selective hybridization between two nucleic acid molecules are well known to those of skill in the art. For example, in some embodiments selective hybridization conditions can be defined as stringent hybridization conditions. For example, stringency of hybridization is controlled by both temperature and salt concentration of either or both of the hybridization and washing steps. For example, the conditions of hybridization to achieve selective hybridization may involve hybridization in high ionic strength solution (6×SSC or 6×SSPE) at a temperature that is about 12-25° C. below the Tm (the melting temperature at which half of the molecules dissociate from their hybridization partners) followed by washing at a combination of temperature and salt concentration chosen so that the washing temperature is about 5° C. to 20° C. below the Tm. The temperature and salt conditions are readily determined empirically in preliminary experiments in which samples of reference DNA immobilized on filters are hybridized to a labeled nucleic acid of interest and then washed under conditions of different stringencies. Hybridization temperatures are typically higher for DNA-RNA and RNA-RNA hybridizations. The conditions can be used as described above to achieve stringency, or as is known in the art. A preferable stringent hybridization condition for a DNA:DNA hybridization can be at about 68° C. (in aqueous solution) in 6×SSC or 6×SSPE followed by washing at 68° C. Stringency of hybridization and washing, if desired, can be reduced accordingly as the degree of complementarity desired is decreased, and further, depending upon the G-C or A-T richness of any area wherein variability is searched for. Likewise, stringency of hybridization and washing, if desired, can be increased accordingly as homology desired is increased, and further, depending upon the G-C or A-T richness of any area wherein high homology is desired, all as known in the art.

Another way to define selective hybridization is by looking at the amount (percentage) of one of the nucleic acids bound to the other nucleic acid. For example, in some embodiments selective hybridization conditions would be when at least about, 60, 65, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100 percent of the limiting nucleic acid is bound to the non-limiting nucleic acid. Typically, the non-limiting primer is in for example, 10 or 100 or 1000 fold excess. This type of assay can be performed at under conditions where both the limiting and non-limiting primer are for example, 10 fold or 100 fold or 1000 fold below their k_(d), or where only one of the nucleic acid molecules is 10 fold or 100 fold or 1000 fold or where one or both nucleic acid molecules are above their k_(d).

Another way to define selective hybridization is by looking at the percentage of primer that gets enzymatically manipulated under conditions where hybridization is required to promote the desired enzymatic manipulation. For example, in some embodiments selective hybridization conditions would be when at least about, 60, 65, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100 percent of the primer is enzymatically manipulated under conditions which promote the enzymatic manipulation, for example if the enzymatic manipulation is DNA extension, then selective hybridization conditions would be when at least about 60, 65, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100 percent of the primer molecules are extended. Conditions also include those suggested by the manufacturer or indicated in the art as being appropriate for the enzyme performing the manipulation.

Just as with homology, it is understood that there are a variety of methods herein disclosed for determining the level of hybridization between two nucleic acid molecules. It is understood that these methods and conditions may provide different percentages of hybridization between two nucleic acid molecules, but unless otherwise indicated meeting the parameters of any of the methods would be sufficient. For example if 80% hybridization was required and as long as hybridization occurs within the required parameters in any one of these methods it is considered disclosed herein.

It is understood that those of skill in the art understand that if a composition or method meets any one of these criteria for determining hybridization either collectively or singly it is a composition or method that is disclosed herein.

Kits

Disclosed herein are kits that are drawn to reagents that can be used in practicing the methods disclosed herein. In particular, he kits can include any reagent or combination of reagents discussed herein or that would be understood to be required or beneficial in the practice of the disclosed methods. For example, the kits could include one or more primers from Tables 7-14 disclosed herein to perform the extension, replication and amplification reactions discussed in certain embodiments of the methods, as well as the buffers and enzymes required to use the primers as intended. The kit can also include other necessary reagents to perform any of the next generation sequencing techniques disclosed herein. In another aspect, the disclosed kits can include one or more of the probes listed in Table 15 in addition to or instead of one or more primers from Table 7-14.

It is understood and herein contemplated that the disclosed kits can comprise at least one primer set to detect the presence of nucleic acid variation in each of KIT, BRAF, KRAS, ALK, and EGFR. For example, the kits can comprise at least one primer or primer set for sequencing at least one of each of the KIT, BRAF, KRAS, ALK, and EGFR exons of Tables 1. In one aspect, the kits can comprise at least one primer or primer set from each of Tables 7-14. Alternatively, the kit can comprise a primer or primer set that will detect one or more of the specific mutations listed in Tables 2-6. Therefore, in one aspect disclosed herein are kits for performing a NGS sequencing reaction on a tissue sample to detect the presence of a mutation conferring kinase inhibitor resistance comprising at least one or more primer or primer set from each of Table 7-14. In another aspect, disclosed herein are kits for performing a NGS sequencing reaction on a tissue sample to detect the presence of a mutation conferring kinase inhibitor resistance comprising at least one or more primer or primer set capable of specifically hybridizing an amplifying any of the mutant sequences of KIT, BRAF, KRAS, ALK, and EGFR present in Tables 2-6.

Additionally, it is understood that the disclosed kits can include such other reagents and material for performing the disclosed methods such as enzymes (e.g., polymerases), buffers, sterile water, and/or reaction tubes. Additionally the kits can also include modified nucleotides, nuclease-resistant nucleotides, and or labeled nucleotides. Additionally, the disclosed kits can include instructions for performing the methods disclosed herein and software for enable the calculation of the presence of a kinase inhibitor mutation (i.e., a mutation in KIT, BRAF, KRAS, EGFR, and/or ALK).

The compositions disclosed herein and the compositions necessary to perform the disclosed methods can be made using any method known to those of skill in the art for that particular reagent or compound unless otherwise specifically noted.

Nucleic Acid Synthesis

The disclosed nucleic acids, such as, the oligonucleotides to be used as primers can be made using standard chemical synthesis methods or can be produced using enzymatic methods or any other known method. Such methods can range from standard enzymatic digestion followed by nucleotide fragment isolation to purely synthetic methods, for example, by the cyanoethylphosphoramidite method using a Milligen or Beckman System 1Plus DNA synthesizer (for example, Model 8700 automated synthesizer of Milligen-Biosearch, Burlington, Mass. or ABI Model 380B).

EXAMPLES

The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how the compounds, compositions, articles, devices and/or methods claimed herein are made and evaluated, and are intended to be purely exemplary and are not intended to limit the disclosure. Efforts have been made to ensure accuracy with respect to numbers (e.g., amounts, temperature, etc.), but some errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, temperature is in ° C. or is at ambient temperature, and pressure is at or near atmospheric.

Example 1 ALK Inhibitor Resistance

Using an in vitro assay known to predict clinically relevant kinase inhibitor-resistance mutations resistance selection studies were performed with XALKORI® and identified a large number ofALK kinase domain point mutations that confer high-level resistance to the Pfizer inhibitor (FIG. 1). In response to the issue of resistance, a number of pharma and biotech companies currently have 2^(nd)-generation ALK small-molecule inhibitors in development.

The need for more comprehensive oncogene profiling in patients with ALK inhibitor resistance was observed in an ALK positive crizotinib resistant cohort of patients that ALK specific kinase mutations accounted for only a third of crizotinib resistance. The larger subset of crizotinib resistant cases indicated that second (co-expression in conjunction with ALK) or separate (complete absence of ALK) oncogenic drivers such as EGFR, BRAF, KRAS or cKIT can relieve the sensitivity to crizotinib and drive oncogenesis. It was also observed in a single case that the complete loss of ALK expression did not correspond to the presence of an identifiable alternate driver indicating the genetic profiling of ALK inhibitor resistance cases should be extended past EGFR, BRAF, KRAS or cKIT expression using more versatile testing platforms. The presence of multiple oncogenes present in a single tumor sample is by no means a new phenomenon as EGFR driven tumors resistant to EGFR tyrosine kinase inhibitors can be driven by secondary MET gene amplification.

Example 2 A Diagnostic Cancer Panel that Employs NGS

Applicants have designed and developed a next generation sequencing panel to amplify and sequence one or more exons within ALK and other oncogenes implicated in driving tumorigenesis in the presence of crizotinib (i.e. ALK, BRAF, EGFR, KIT and KRAS. See Table 1 for an overarching description of the exons targeted for sequencing in the panel and Tables 2-6 for a more detailed list of each mutation detected by the Insight ALK resistance ID™ panel. Primer sequences used to amplify each gene segment are depicted in Tables 7-14.

TABLE 1 Exons That Are Covered Gene Exon KRAS 1 KRAS 2 EGFR 18 EGFR 19 EGFR 20 EGFR 21 EGFR 22 ALK 21 ALK 22 ALK 23 ALK 24 ALK 25 KIT 8 KIT 9 KIT 10 KIT 11 KIT 12 KIT 13 KIT 17 BRAF 10 BRAF 11 BRAF 13 BRAF 14 BRAF 15

Example 3 Targeted Next Generation Sequencing Insight ALK Resistance ID™

Polymerase chain reaction is used to create amplicons that span the exonic regions mentioned above. The design described here is agnostic to the NGS platform used to perform the actual sequencing, and thus multiple PCR strategies can match the size of the PCR fragments to the read-length of the sequencing platform being employed. The PCR amplification can be done in a single-tube as a multiple reaction where all targets are covered at once. In the case of low coverage or ambiguous results, a single-plex PCR can be performed as a confirmatory step to ensure accurate mutation calling. This is also true in the case of highly-degraded samples where the template DNA has fragmented and large-amplicons cannot be extracted from the DNA that remains. See Tables 7-14 for a full list of the primers that have been designed and the general size of fragments each set produces. There are a large number of primers in the list to ensure that there is flexibility to run various multiplex PCR reactions where there is very little sequence overlap in the primers, which can lead to dimerization, and allow melting temperatures of all the oligos in a particular reaction to be matched. The amplification parameters of each PCR reaction consist of 95° C. 15-min heat denaturation phase followed by 40 cycles of denaturation at 95° C. for 15 sec and 55° C. annealing for 30 sec and 72° C. extension for 1 min and finally a 72° C. final extension step for 5 minutes. At the end of the PCR step a diverse set of fragments that cover the exons of interest can be synthesized. The fragments can then be adapted for sequencing on any commercially available NGS platform. Since there is a very wide range of read-lengths that the different NGS instruments produce, from as low as 35 bases to as high as 1500 and expectations of 100 kb read length in the near future, the Insight ALK resistance ID™ is designed to be able to produce fragments as short as 150 bases to as high as 5kb. This ensures for efficient sequencing where the size of each amplicon can be matched to the output of long-read and middle-read technologies (150-1000 bases) or have large enough fragments (5kb) that can be effectively sheared, either sonically or enzymatically, to be compatible with short-read sequencers (<150 bases).

The ALK resistance ID™ takes advantage of the very high-throughput offered by modern sequencers to cover the regions of interest at very high coverage (depth>5,000×) and thus enable the detection of rare variants only present in the sample at a frequency of 1% or less. The sequence reads that are generated can be compared to a reference sequence examined for the presence of any of the mutations listed in Tables 2-6.

TABLE 2 ALK Mutations That Are Covered Amino Acid Mutation Nucleotide Mutation p.V1471fs*45 c.4409_4422delCCGTGGAAGGGGGA p.Y1584Y c.4752C > T p.T1597T c.4791T > A p.L1062I c.3185A > T p.T1087I c.3260C > T p.D1091N c.3271G > A p.G1128A c.3383G > C p.M1166R c.3497T > G p.A1168P c.3502C > G p.I1171N c.3512T > A p.F1174I c.3520T > A p.F1174L c.3522C > A p.R1192P c.3575G > C p.F1245C c.3734T > G p.F1245V c.3733T > G p.F1245L c.3735C > G p.F1245I c.3733T > A p.I1250T c.3749T > C p.R1275Q c.3824G > A

TABLE 3 EGFR Mutations That Are Covered Amino Acid Mutation Nucleotide Mutation p.L747_T751 > S c.2240_2251del12 p.L861Q c.2582T > A p.L747_E749del c.2239_2247del9 p.E746_S752 > D c.2238_2255del18 p.E746_A750del c.2235_2249del15 p.L858R c.2573T > G p.E746_A750del c.2236_2250del15 p.R776C c.2326C > T p.H835L c.2504A > T p.G719A c.2156G > C p.T790M c.2369C > T p.S768I c.2303G > T p.V769L c.2305G > T p.G719S c.2155G > A p.G719C c.2155G > T p.L747_T751del c.2239_2253del15 p.L747_S752del c.2239_2256del18 p.S752_I759del c.2254_2277del24 p.P753S c.2257C > T p.L858M c.2572C > A p.E746_S752 > A c.2237_2254del18 p.L747_T751del c.2240_2254del15 p.L747_P753 > S c.2240_2257del18 p.E709V c.2126A > T p.I715S c.2144T > G p.S720F c.2159C > T p.L861R c.2582T > G p.V769_D770insASV c.2307_2308ins9 p.H773_V774insH c.2319_2320insCAC p.D770_N771insG c.2310_2311insGGT p.V769_D770insCV c.2307_2308insTGCGTG p.H773_V774insPH c.2319_2320insCCCCAC p.H773_V774insNPH c.2319_2320ins9 p.L747_A750 > P c.2239_2248TTAAGAGAAG > C p.L747_T751 > P c.2239_2251 > C p.E746_S752 > V c.2237_2255 > T p.E746_S752 > I c.2235_2255 > AAT p.E746_T751 > V c.2237_2252 > T p.L747_P753 > Q c.2239_2258 > CA p.H773 > NPY c.2317_2317C > AACCCCT p.V774_C775insHV c.2322_2322G > CCACGTG p.L747_S752 > Q c.2239_2256 > CAA p.E746_T751 > I c.2229_2252 > AATTAAGA p.T751_I759 > S c.2252_2275 > G p.E746_A750 > RP c.2236_2248 > AGAC p.E746_T751 > VA c.2237_2253 > TTGCT p.L747_T751 > Q c.2238_2252 > GCA p.L747_T751 > Q c.2239_2252 > CA p.L747_S752 > QH c.2238_2255 > GCAACA p.L747_A750 > P c.2238_2248 > GC p.I744_K745insKIPVAI c.2231_2232ins18 p.D761_E762insEAFQ c.2283_2284ins12 p.A767_S768insTLA c.2302_2303ins9 p.V769_D770insASV c.2308_2309ins9 p.D770 > GY c.2308_2309insGTT p.E709H c.2125_2127GAA > CAT p.L858R c.2573_2574TG > GT p.A859T c.2575G > A p.E746_T751 > A c.2237_2251del15 p.Y727C c.2180A > G p.V851I c.2551G > A p.E746_T751del c.2236_2253del18 p.D770_N771 > AGG c.2309_2312ACAA > CTGGTGG p.G857R c.2569G > A p.L858R c.2573.T > G p.E746_A750del c.2235_2249del15 p.E746_A750 > QP c.2236_2248 > CAAC p.G810D c.2429G > A p.E709K c.2125G > A p.D770_N771insN c.2310_2311insAAC p.D770_N771insG c.2310_2311insGGC p.H773L c.2318A > T p.V774M c.2320G > A p.G779F c.2335_2336GG > TT p.A871G c.2612C > G p.E709G c.2126A > G p.L861Q c.2582T > A p.L730F c.2188C > T p.P733L c.2198C > T p.G735S c.2203G > A p.V742A c.2225T > C p.E746K c.2236G > A p.T751I c.2252C > T p.S752Y c.2255C > A p.H850N c.2548C > A p.D761N c.2281G > A p.S784F c.2351C > T p.L792P c.2375T > C p.L798F c.2392C > T p.G810S c.2428G > A p.N826S c.2477A > G p.T847I c.2540C > T p.V851A c.2552T > C p.I853T c.2558T > C p.A864T c.2590G > A p.E866K c.2596G > A p.G873E c.2618G > A p.E746_P753 > LS c.2236_2257 > CTCT p.V819V c.2457G > A p.Y764Y c.2292C > T p.L833V c.2497T > G p.V769M c.2305G > A p.L838V c.2512C > G p.E709A c.2126A > C p.D770_N771insSVD c.2311_2312ins9 p.A839T c.2515G > A p.H773R c.2318A > G p.P772P c.2316C > T p.E746_T751 > A c.2235_2251 > AG p.E746_A750 > IP c.2235_2248 > AATTC p.E746_T751 > I c.2235_2252 > AAT p.E746_T751 > IP c.2235_2251 > AATTC p.L858R c.2572_2573CT > AG p.N771_P772 > SVDNR c.2312_2315ACCC > GCGTGGACAACCG p.D770_P772 > ASVDNR c.2308_2315GACAACCC > CCAGCGTGGATAACCG p.S752_I759del c.2253_2276del24 p.E746_A750 > QP c.2236_2248 > CAAC p.V769_D770insASV c.2309_2310AC > CCAGCGTGGAT p.M766_A767insAI c.2298_2299insGCCATA p.G724S c.2170G > A p.D770N c.2308G > A p.T783I c.2348C > T p.G863D c.2588G > A p.V897I c.2689G > A p.K745R c.2234A > G p.P741L c.2222C > T p.E734K c.2200G > A p.E746del c.2234_2236delAGG p.E746_T751 > VP c.2237_2251 > TTC p.Q787R c.2360A > G p.V834L c.2500G > T p.A755A c.2265C > T p.G719D c.2156G > A p.E746_S752 > V c.2237_2256 > TC p.E746_P753 > VS c.2237_2257 > TCT p.E746_A750 > DP c.2238_2249 > TCC p.V769_D770insGSV c.2308_2309ins9 p.V769_D770insGVV c.2308_2309ins9 p.N771 > GF c.2311_2312AA > GGGTT p.V774_C775insHV c.2321_2322insCCACGT p.G719C c.2154_2155GG > TT p.L747_R748 > FP c.2241_2244AAGA > CCCG p.E872 c.2614G > T p.G873G c.2619A > T p.P753P c.2259G > A p.G719fs*29 c.2156delG p.L747_K754 > ST c.2240_2261 > CGAC p.S768_V769insVAS c.2303_2304ins9 p.V769_D770insDNV c.2307_2308ins9 p.D770_N771insAPW c.2310_2311ins9 p.N771_P772insN c.2313_2314insAAC p.G796S c.2386G > A p.E804G c.2411A > G p.R841K c.2522G > A p.V834M c.2500G > A p.D761Y c.2281G > T p.R776H c.2327G > A p.L778L c.2334G > T p.G779C c.2335G > T p.P848L c.2543C > T p.L747_T751 > P c.2238_2251 > GC p.T751_I759 > S c.2251_2277 > TCT p.N771 > TH c.2311_2312insCAC p.H773_V774insPH c.2318_2319insCCCCCA p.V774_C775insHV c.2322_2323insCACGTG p.L862P c.2585T > C p.S784Y c.2351C > A p.F795S c.2384T > C p.F795S c.2384T > C p.Y813C c.2438A > G p.Y801H c.2401T > C p.C775Y c.2324G > A p.D770_N771insDG c.2308_2309insACGGCG p.T751_I759 > REA c.2252_2277 > GAGAAGCG p.L777Q c.2330T > A p.G721S c.2161G > A p.G721D c.2162G > A p.K754K c.2262A > G p.E746_T751 > Q c.2236_2253 > CAA p.L747_T751del c.2238_2252del15 p.L747_T751 > A c.2239_2253 > GCT p.C818Y c.2453G > A p.I759N c.2276T > A p.T751_E758del c.2250_2273del24 p.L747P c.2239_2240TT > CC p.L858K c.2572_2573CT > AA p.P753_I759del c.2257_2277del21 p.T751_I759 > N c.2252_2277 > AT p.G863G c.2589T > G p.N771 > SH c.2311_2312insGTC p.D770fs*61 c.2309_2310ins14 p.E829E c.2487G > A p.R831C c.2491C > T p.R831C c.2491C > T p.L861V c.2581C > G p.E746_T751del c.2235_2252del18 p.L747_K754del c.2239_2262del24 p.L838P c.2513T > C p.K757 > NK c.2270_2271insCAA p.G779S c.2335G > A p.V774L c.2320G > T p.L815L c.2445C > T p.E758D c.2274A > C p.K875R c.2624A > G p.A864E c.2591C > A p.Y869C c.2606A > G p.K745_E749del c.2233_2247del15 p.F723F c.2169C > T p.L858L c.2572C > T p.A859fs*38 c.2575_2576insG p.N756S c.2267A > G p.V845A c.2534T > C p.F856S c.2567T > C p.G874S c.2620G > A p.A750_E758del c.2247_2273del27 p.A750_E758 > P c.2248_2273 > CC p.L747_5752 > Q c.2238_2256 > GCAA p.A859_L883 > V c.2576_2647del72 p.I744_K745insKIPVAI c.2232_2233ins18 p.K745_E746insVPVAIK c.2236_2237ins18 p.A767V c.2300C > T p.N842D c.2524A > G p.A743T c.2227G > A p.L747S c.2240T > C p.K860I c.2579A > T p.A750_K754del c.2246_2260del15 p.D770_N771insMATP c.2311_2312ins12 p.A763_Y764insFQEA c.2290_2291ins12 p.D761G c.2282A > G p.V786M c.2356G > A p.G796A c.2387G > C p.K728 c.2182A > T p.R832C c.2494C > T p.G721A c.2162G > C p.I744V c.2230A > G p.S784P c.2350T > C p.R832L c.2495G > T p.V802F c.2404G > T p.E746_E749del c.2235_2246del12 p.T854A c.2560A > G p.E884K c.2650G > A p.F712S c.2135T > C p.I744M c.2232C > G p.V765M c.2293G > A p.R836C c.2506C > T p.A871T c.2611G > A p.D855G c.2564A > G p.E868G c.2603A > G p.L798H c.2393T > A p.K806E c.2416A > G p.L814P c.2441T > C p.E746_A750 > VP c.2237_2250 > TCCCT p.V769_D770insMASVD c.2307_2308ins15 p.F723S c.2168T > C p.T785N c.2354C > A p.V845M c.2533G > A p.M766T c.2297T > C p.S752P c.2254T > C p.T725T c.2175G > A p.D855N c.2563G > A p.L858Q c.2573T > A p.H870R c.2609A > G p.F712L c.2134T > C p.I821T c.2462T > C p.V834A c.2501T > C p.L718P c.2153T > C p.D770_N771insNPH c.2310_2311insAACCCCCAC p.D770_N771insGL c.2310_2311insGGGTTA p.D770_N771insSVD c.2311_2312insGCGTGGACA p.P772_H773insTHP c.2315_2316insGACACACCC p.S720T c.2158T > A p.E746V c.2237_2238AA > TT p.E746_P753 > VQ c.2237_2258 > TTCA p.E709_T710 > D c.2127_2129delAAC p.E746_T751 > IP c.2236_2253 > ATTCCT p.L747_T751 > Q c.2239_2253 > CAA p.H773_V774insGNPH c.2320_2321ins12 p.I732T c.2195T > C p.N756Y c.2266A > T p.L844P c.2531T > C p.I740T c.2219T > C p.E746_T751 > VP c.2237_2253 > TTCCT p.W731L c.2192G > T p.E734Q c.2200G > C p.T785A c.2353A > G p.C797Y c.2390G > A p.R831H c.2492G > A p.N771 > GY c.2311_2311A > GGTT p.P733S c.2197C > T p.R748I c.2243G > T p.Q849R c.2546A > G p.E746_T751 > VA c.2237_2251 > TGG p.E868D c.2604A > T p.S720S c.2160C > A p.T725A c.2173A > G p.R836S c.2506C > A p.I744I c.2232C > A p.E866G c.2597A > G p.I853I c.2559C > T p.K708E c.2122A > G p.G824G c.2472C > A p.F712F c.2136C > T p.Y827Y c.2481C > T p.T725M c.2174C > T p.T725M c.2174C > T p.K852N c.2556G > T p.A722V c.2165C > T p.E711K c.2131G > A p.T785I c.2354C > T p.D800N c.2398G > A p.E872G c.2615A > G p.E829K c.2485G > A p.E829K c.2485G > A p.H870Y c.2608C > T p.H870Y c.2608C > T p.D770_N771insSVD c.2310_2311ins9 p.S768_V769 > IL c.2303_2305GCG > TCT p.D770_N771insGD c.2310_2311insGGGGAC p.E709_T710 > A c.2126_2128delAAA p.E746_S752 > V c.2235_2255 > GGT p.I744_A750 > VK c.2230_2249 > GTCAA p.L747_K754 > N c.2239_2264 > GCCAA p.I740_P741insPVAIKI c.2219_2220ins18 p.R836R c.2508C > T p.V843I c.2527G > A p.K754R c.2261A > G p.A840T c.2518G > A p.K754E c.2260A > G p.A859D c.2576C > A p.Y801C c.2402A > G p.I744T c.2231T > C p.T854I c.2561C > T p.G863S c.2587G > A p.H850R c.2549A > G p.K754A c.2260_2261AA > GC p.D807N c.2419G > A p.S720P c.2158T > C p.K757M c.2270A > T p.L862Q c.2585T > A p.T751_I759 > N c.2252_2276 > A p.P772R c.2315C > G p.A839V c.2516C > T p.K716R c.2147A > G p.H773_V774insQ c.2319_2320insCAG p.E711V c.2132_2133AA > TT p.T710A c.2128A > G p.K714N c.2142G > C p.V717A c.2150T > C p.G729E c.2186G > A p.I744_E749 > LKR c.2230_2247 > CTTAAGAGA p.E746_T751 > L c.2236_2253 > CTA p.E746_S752 > I c.2236_2255 > AT p.E746_S752del c.2236_2256del21 p.E746_S752 > I c.2236_2256 > ATC p.E746_P753 > IS c.2236_2259 > ATCTCG p.E746_T751 > V c.2237_2253 > TA p.E746_T751 > V c.2237_2253 > TC p.L747_A750 > P c.2239_2250 > CCA p.L747_S752 > QH c.2239_2256 > CAACAT p.L747_P753 > S c.2239_2257 > T p.R748K c.2243G > A p.E749G c.2246A > G p.T751_I759 > S c.2251_2277 > TCC p.P772_H773insHV c.2316_2317insCACGTG p.G779D c.2336G > A p.V802A c.2405T > C p.L833W c.2498T > G p.D837G c.2510A > G p.L844V c.2530C > G p.T751_I759del c.2252_2275del24 p.V765G c.2294T > G p.G796D c.2387G > A p.R836H c.2507G > A p.K757R c.2270A > G p.E872K c.2614G > A p.L858L c.2574G > A p.I780S c.2339T > G p.T785P c.2353A > C p.Y801fs*1 c.2402_2403insG p.L858R c.2573_2574TG > GA p.N771_P772insRH c.2311_2312insACCGGC p.H850Y c.2548C > T p.E868K c.2602G > A p.I780T c.2339T > C p.E866D c.2598G > T p.L833F c.2499G > T p.A864V c.2591C > T p.K745_A750del c.2232_2249del18 p.P794H c.2381C > A p.E804K c.2410G > A p.G857E c.2570G > A

TABLE 4 KIT Mutations That Are Covered Amino Acid Mutation Nucleotide Mutation p.(550_592)ins7 c.(1648_1774)ins21 p.C443Y c.1328G > A p.P456S c.1366C > T p.L462L c.1384C > T p.P468P c.1404G > A p.F469L c.1405T > C p.L472L c.1416A > G p.S476I c.1427G > T p.D479fs*2 c.1434_1462del29 p.S480fs*47 c.1439delC p.N486D c.1456A > G p.V489I c.1465G > A p.V489A c.1466T > C p.E490G c.1469A > G p.E490_F504 > DHIVVSLTF c.1470_1512 > CCACATCGTTGTAAGCCTTACATTC p.N495I c.1484A > T p.N495I c.1484A > T p.D496V c.1487A > T p.V50M c.148G > A p.V497V c.1491G > A p.G498D c.1493G > A p.K499K c.1497G > A p.A502_Y503insSA c.1504_1505insCTTCTG p.A502_Y503insSA c.1505_1506insTTCTGC p.Y503_F504insSA c.1507_1508insCTGCCT p.A502_Y503insFA c.1507_1508insTTGCCT p.Y503_F504insAY c.1509_1510insGCCTAT p.F504L c.1510T > C p.N505H c.1513A > C p.F506L c.1516T > C p.F506_A507insAYFNF c.1518_1519ins15 p.F508_K509insNFAF c.1524_1525ins12 p.K509I c.1526A > T p.G510del c.1528_1530delGGT p.N512D c.1534A > G p.V530I c.1588G > A p.I531_V532insGF c.1593_1594insGGGTTC p.M541L c.1621A > C p.K546K c.1638A > G p.Q549_V555 > I c.1645_1663 > A p.K550_P551del c.1648_1653delAAACCC p.K550_E554del c.1648_1662del15 p.K550_V555del c.1648_1665del18 p.K550_V555del c.1648_1665del18 p.K550_Q556 > II c.1648_1668 > ATTATT p.K550_W557del c.1648_1671del24 p.K550fs*6 c.1648_1672del25 p.K550_K558del c.1648_1674del27 p.K550_V559del c.1648_1677del30 p.K550_V555 > I c.1649_1663del15 p.K550R c.1649A > G p.K550I c.1649A > T p.K550_V555 > KTL c.1650_1663 > AACCC p.P551_K558del c.1650_1673del24 p.K550N c.1650A > C p.P551del c.1651_1653delCCC p.P551_E554del c.1651_1662del12 p.P551_V555del c.1651_1665del15 p.P551_Q556del c.1651_1668del18 p.P551T c.1651C > A p.P551S c.1651C > T p.P551_M552 > L c.1652_1654delCCA p.P551_E554 > H c.1652_1662 > AY p.P551_V555 > L c.1652_1663del12 p.P551_V559del > L c.1652_1678del27 p.P551L c.1652C > T p.M552_Y553del c.1653_1658delCATGTA p.M552_Q556> c.1653_1667 > TCT p.M552_W557del c.1653_1670del18 p.M552_Y553del c.1654_1659delATGTAT p.M552_E554del c.1654_1662del9 p.M552_V555del c.1654_1665del12 p.M552_Q556del c.1654_1668del15 p.M552_W557del c.1654_1671del18 p.M552_K558del c.1654_1674del21 p.M552_D572del c.1654_1716del63 p.M552L c.1654A > C p.M552L c.1654A > C p.M552_Y553 > N c.1655_1657delTGT p.M552_E554 > K c.1655_1660delTGTATG p.M552_V555 > I c.1655_1663del9 p.M552_Q556 > K c.1655_1666del12 p.M552_W557 > R c.1655_1669del15 p.M552_W557del c.1655_1672del18 p.M552_K558 > T c.1655_1674 > CN p.M552_E561 > K c.1655_1681del27 p.M552_T574 > TESA c.1655_1720 > CAGAATCAG p.M552K c.1655T > A p.M552T c.1655T > C p.Y553_W557del c.1656_1670del15 p.Y553_K558> c.1656_1673del18 p.Y553V c.1657_1658TA > GT p.Y553_Q556del c.1657_1668del12 p.Y553_W557del c.1657_1671del15 p.Y553_K558del c.1657_1674del18 p.Y553_V559 > E c.1657_1677 > GAA p.Y553_V559del c.1657_1677del21 p.Y553N c.1657T > A p.Y553_T574 > S c.1658_1720del63 p.E554_K558del c.1660_1674del15 p.E554_E562del c.1660_1686del27 p.E554_N564del c.1660_1692del33 p.E554_I571del c.1660_1713del54 p.E554_D572del c.1660_1716del57 p.E554K c.1660G > A p.E554K c.1660G > A p.E554_K558del c.1661_1675del15 p.E554G c.1661A > G p.V555_E562del c.1662_1685del24 p.E554D c.1662A > T p.V555_Q556del c.1663_1668delGTACAG p.V555_K558del c.1663_1674del12 p.V555_V559del c.1663_1677del15 p.V555_V560del c.1663_1680del18 p.V555_I563del c.1663_1689del27 p.V555_G565del c.1663_1695del33 p.V555_Y570del c.1663_1710del48 p.V555_I571del c.1663_1713del51 p.V555_P573del c.1663_1719del57 p.V555I c.1663G > A p.V555_N566 > D c.1664_1696del33 p.Q556_V559del c.1665_1676del12 p.V555_V560 > V c.1665_1679del15 p.Q556_N566 > SNNLQLY c.1665_1696 > TTCCAACAACCTTCCACTGT p.Q556_D572del c.1665_1716 > T p.Q556_W557del c.1666_1671delCAGTGG p.Q556_V559del c.1666_1677del12 p.Q556_V560 > F c.1666_1678 > T p.Q556_V560 > TTF c.1666_1680 > ACAACCTTC p.Q556_V560del c.1666_1680del15 p.Q556_E561 > HH c.1666_1683 > CATCAT p.Q556_E561del c.1666_1683del18 p.Q556_D572 > PS c.1666_1716 > CCATCC p.Q556_P573del c.1666_1719del54 p.Q556_T574del c.1666_1722del57 p.Q556_L576del c.1666_1728del63 p.Q556_W557 > R c.1667_1669delAGT p.W557_K558del c.1667_1672delAGTGGA p.Q556_K558 > R c.1667_1673AGTGGAA > G p.W557_E561del c.1667_1681del15 p.Q556R c.1667A > G p.W557_K558del c.1668_1673delGTGGAA p.Q556_K558 > HPCR c.1668_1673GTGGAA > CCCCTGCAG p.Q556_K558 > H c.1668_1674GTGGAAG > Y p.Q556_V559 > H c.1668_1676del9 p.Q556_V559 > HT c.1668_1677GTGGAAGGTT > TACT p.Q556_V560 > HNLQLY c.1668_1679 > CAACCTTCCACTGTA p.Q556_V560 > H c.1668_1679del12 p.W557_I571del c.1668_1712del45 p.Q556_D572 > H c.1668_1715del48 p.W557_Q575del c.1668_1724del57 p.W557del c.1669_1671delTGG p.W557_K558 > E c.1669_1672TGGA > G p.W557_K558del c.1669_1674delTGGAAG p.W557_K558 > S c.1669_1674TGGAAG > C p.W557_V559 > I c.1669_1675TGGAAGG > A p.W557_V559del c.1669_1677del9 p.W557_V560del c.1669_1680del12 p.W557_E561del c.1669_1683del15 p.W557_E562del c.1669_1686del18 p.W557_Q575del c.1669_1725del57 p.W557R c.1669T > A p.W557R c.1669T > C p.W557G c.1669T > G p.W557_K558 > SS c.1670_1673GGAA > CTTC p.W557_K558 > FP c.1670_1674GGAAG > TTCCT p.W557_V559 > F c.1670_1675delGGAAGG p.W557_V560 > F c.1670_1678del9 p.W557_P573 > S c.1670_1717del48 p.W557S c.1670G > C p.W557_K558 > CT c.1671_1673GAA > CAC p.W557_K558 > CP c.1671_1673GAA > TCC p.W557_K558 > C c.1671_1674GAAG > C p.W557_V559 > C c.1671_1676delGAAGGT p.W557_V560 > C c.1671_1679del9 p.W557 c.1671G > A p.W557C c.1671G > T p.K558_V559 > SS c.1672_1676AAGGT > TCTTC p.K558_V559del c.1672_1677delAAGGTT p.K558_V560del c.1672_1680del9 p.K558_E562del c.1672_1686del15 p.K558_N564del c.1672_1692del21 p.K558_G565del c.1672_1695del24 p.K558_D572del c.1672_1716del45 p.K558_Q575del c.1672_1725del54 p.K558E c.1672A > G p.K558* c.1672A > T p.K558 > NP c.1673_1674insTCC p.K558_V560 > I c.1673_1678delAGGTTG p.K558_V560 > M c.1673_1680AGGTTGTT > TG p.K558_E562del c.1673_1687del15 p.K558_G565 > R c.1673_1693del21 p.K558R c.1673A > G p.K558 > NP c.1674_1674G > TCCT p.K558_V559 > N c.1674_1676delGGT p.K558_V560 > N c.1674_1679delGGTTGT p.K558_Y570 > N c.1674_1709del36 p.K558_L576 > NV c.1674_1726 > CG p.K558K c.1674G > A p.K558N c.1674G > C p.K558N c.1674G > Y p.V559del c.1675_1677delGTT p.V559K c.1675_1677GTT > AAG p.V559_V560del c.1675_1680delGTTGTT p.V559_E561del c.1675_1683del9 p.V559_G565del c.1675_1695del21 p.V559_I571del c.1675_1713del39 p.V559_L576del c.1675_1728del54 p.V559I c.1675G > A p.V559_E561del c.1676_1684del9 p.V559_E562del c.1676_1687del12 p.V559_P573 > A c.1676_1717del42 p.V559D c.1676T > A p.V559A c.1676T > C p.V559G c.1676T > G p.V560del c.1678_1680delGTT p.V560_L576del c.1678_1728del51 p.V560E c.1679_1680TT > AG p.V560E c.1679_1680TT > AR p.V560del c.1679_1681delTTG p.V560_I571del c.1679_1714del36 p.V560D c.1679T > A p.V560A c.1679T > C p.V560G c.1679T > G p.E561del c.1680_1682delTGA p.V560V c.1680T > G p.E561del c.1681_1683delGAG p.E561_P577del c.1681_1731del51 p.E561K c.1681G > A p.E561G c.1682A > G p.E561E c.1683G > A p.E562_P573del c.1684_1719del36 p.E562K c.1684G > A p.E562V c.1685A > T p.E562_V569 > D c.1686_1706del21 p.I563_D572del c.1687_1716del30 p.I563_L576del c.1687_1728del42 p.I563V c.1687A > G p.N564_T574del c.1690_1722del33 p.N564_L576del c.1690_1728del39 p.N564_P577del c.1690_1731del42 p.N564_Y578del c.1690_1734del45 p.N564H c.1690A > C p.N564_P573 > TS c.1691_1717 > CCT p.N564_P573 > T c.1691_1717del27 p.N564S c.1691A > G p.N564K c.1692T > G p.G565R c.1693G > A p.G565E c.1694G > A p.G565V c.1694G > T p.N566D c.1696A > G p.N566S c.1697A > G p.N567_L576 > E c.1698_1728 > CGAA p.N566N c.1698C > T p.N567_P573del c.1699_1719del21 p.N567H c.1699A > C p.N567K c.1701T > A p.Y568_T574del c.1702_1722del21 p.Y568D c.1702T > G p.Y568_L576 > CV c.1703_1726 > GTG p.Y568S c.1703A > C p.Y568C c.1703A > G p.Y568Y c.1704T > C p.V569_L576del c.1705_?del? p.V569_D572del c.1705_1716del12 p.V569_Q575del c.1705_1725del21 p.V569_L576del c.1705_1728del24 p.V569I c.1705G > A p.Y570_L576delYIDPTQL c.1706_1726del21 p.V569_L576 > G c.1706_1727 > G p.V569A c.1706T > C p.V569G c.1706T > G p.Y570_L576del c.1708_1728del21 p.Y570D c.1708T > G p.Y570* c.1710C > A p.I571_L576del c.1711_1728del18 p.I571_N587del c.1712_1762del51 p.I571R c.1712T > G p.I571M c.1713A > G p.571_572 > GE c.1714_1715insGGGAAG p.D572N c.1714G > A p.D572Y c.1714G > T p.D572A c.1715A > C p.D572D c.1716C > T p.P573L c.1718C > T p.P573_T574insYIDP c.1719_1720ins12 p.T574A c.1720A > G p.T574_Q575ins12 c.1721_1722ins36 p.T574I c.1721C > T p.Q575del c.1723_1725delCAA p.Q575_P577 > T c.1723_1731CAACTTCCT > ACA p.L576del c.1726_1728delCTT p.L576F c.1726C > T p.L576del c.1727_1729delTTC p.L576P c.1727T > C p.L576_P577insQL c.1728_1729insCAACTT p.P577_Y578del c.1729_1734delCCTTAT p.P577S c.1729C > T p.P577_D579del c.1730_1738del9 p.P577H c.1730C > A p.P577L c.1730C > T p.D579del c.1735_1737delGAT p.D579_H580insPTQLPYD c.1737_1738ins21 p.D579_H580insSYD c.1737_1738ins9 p.H580del c.1737_1739delTCA p.H580Y c.1738C > T p.H580_K581insHPYD c.1739_1740ins12 p.H580_K581insPYDH c.1740_1741ins12 p.H580_K581insPTQLPYDH c.1740_1741ins24 p.H580_K581insIDPTQLPYDH c.1740_1741ins30 p.H580_K581insYDH c.1740_1741ins9 p.K581R c.1742A > G p.W582* c.1745G > A p.W582* c.1746G > A p.E583_F584insPYDHKWE c.1748_1749ins21 p.E583G c.1748A > G p.F584L c.1750T > C p.F584S c.1751T > C p.F584_P585insLPYDHKWEF c.1752_1753ins27 p.F584_P585ins13 c.1752_1753ins39 p.F584_P585ins15 c.1752_1753ins45 p.P585_R586insYDHKWEFP c.1754_1755ins24 p.P585_R586ins12 c.1754_1755ins36 p.P585_R586insLPYDHKWEFP c.1755_1756ins30 p.P585_R586ins13 c.1755_1756ins39 p.P585_R586ins14 c.1755_1756ins42 p.P585_R586ins17 c.1755_1756ins51 p.P585P c.1755C > T p.N587_R588ins15 c.1761_1762ins45 p.N587N c.1761C > T p.R588_L589ins17 c.1764_1765ins51 p.S590N c.1769G > A p.F591L c.1771T > C p.F591_G592ins21 c.1773_1774ins63 p.G592_K593ins16 c.1774_1775ins48 P.? c.1774 + 3C > T p.G592_K593ins21 c.1775_1776ins63 p.T594I c.1781C > T p.A599T c.1795G > A p.P627L c.1880C > T p.T632I c.1895C > T p.E633G c.1898A > G p.R634R c.1902G > A p.E635G c.1904A > G p.A636V c.1907C > T p.L637F c.1909C > T p.S639P c.1915T > C p.K642Q c.1924A > C p.K642E c.1924A > G p.V643A c.1928T > C p.S645N c.1934G > A p.L647F c.1939C > T p.L647P c.1940T > C p.G648S c.1942G > A p.N649_H650insN c.1947_1948insAAT p.I653T c.1958T > C p.V654A c.1961T > C p.N655K c.1965T > G p.G663V c.1988G > T p.G664R c.1990G > A p.T670E c.2008_2009AC > GA p.T670I c.2009C > T p.L682fs*1 c.2045delT p.S692L c.2075C > T p.E695K c.2083G > A p.H697Y c.2089C > T p.H697fs*28 c.2089delC p.R815_D816insVI c.2445_2446insGTCATA p.D816I c.2446_2447GA > AT p.D816F c.2446_2447GA > TT p.D816N c.2446G > A p.D816H c.2446G > C p.D816Y c.2446G > T p.D816 > GP c.2447_2448AC > GGCCA p.D816 > VVA c.2447_2448AC > TCGTTGCA p.D816A c.2447A > C p.D816G c.2447A > G p.D816V c.2447A > T p.D816E c.2448C > G p.D816E c.2448C > G p.I817V c.2449A > G p.I817T c.2450T > C p.K818R c.2453A > G p.K818K c.2454G > A p.N819Y c.2455A > T p.D820N c.2458G > A p.D820H c.2458G > C p.D820H c.2458G > C p.D820Y c.2458G > T p.D820Y c.2458G > T p.D820A c.2459A > C p.D820G c.2459A > G p.D820V c.2459A > T p.D820E c.2460T > A p.D820E c.2460T > G p.N822H c.2464A > C p.N822Y c.2464A > T p.N822Y c.2464A > T p.N822S c.2465A > G p.N822K c.2466T > A p.N822N c.2466T > C p.N822K c.2466T > G p.N822K c.2466T > R p.Y823N c.2467T > A p.Y823D c.2467T > G p.Y823C c.2468A > G p.V825I c.2473G > A p.V825A c.2474T > C p.A829P c.2485G > C p.A829V c.2486C > T p.R830* c.2488C > T p.R830* c.2488C > T p.L831P c.2492T > C p.V833L c.2497G > C p.V833V c.2499G > T p.E839K c.2515G > A p.C844Y c.2531G > A p.Y846H c.2536T > C p.F848L c.2542T > C p.E849* c.2545G > T p.W853* c.2558G > A p.S854P c.2560T > C p.L859P c.2576T > C p.L859L c.2577T > G p.E861E c.2583G > A p.L862L c.2586G > C

TABLE 5 KRAS Mutations That Are Covered Amino Acid Mutation Nucleotide Mutation p.V9V c.27T > C p.A11P c.31G > C p.A11V c.32C > T p.G12F c.34_35GG > TT p.G12C c.34_36GGT > TGC p.G12L c.34_35GG > CT p.G12L c.34_35GG > CT p.G12V c.35_36GT > TC p.G12C c.34G > T p.G12S c.34G > A p.G12R c.34G > C p.G12E c.35_36GT > AA p.G12V c.35G > T p.G12D c.35G > A p.G12A c.35G > C p.G12G c.36T > C p.G13C c.37G > T p.G13S c.37G > A p.G13R c.37G > C p.G13D c.38G > A p.G13A c.38G > C p.G13V c.38G > T p.A18T c.52G > A p.A18D c.53C > A p.Q61K c.181C > A p.Q61E c.181C > G p.Q61P c.182A > C p.Q61R c.182A > G p.Q61L c.182A > T p.Q61H c.183A > C p.Q61H c.183A > T p.D69fs*4 c.205delG p.G12fs*3 c.35delG p.G13V c.38_39GC > TT p.V14I c.40G > A p.Q61K c.180_181TC > CA

TABLE 6 BRAF Mutations That Are Covered Amino Acid Mutation cDNA Nucleotide Mutation p.G30D c.89G > A p.M53T c.158T > C p.S102F c.305C > T p.S129L c.386C > T p.R146W c.436C > T p.I156I c.468C > T p.R178* c.532C > T p.A184T c.550G > A p.Y198H c.592T > C p.Q201H c.603G > T p.K205Q c.613A > C p.F247L c.741T > G p.Q257H c.771G > T p.G258V c.773G > T p.H298Y c.892C > T p.I300V c.898A > G p.A305V c.914C > T p.E309* c.925G > T p.T310I c.929C > T p.S323S c.969G > A p.I326V c.976A > G p.I326T c.977T > C p.F357S c.1070T > C p.G358G c.1074G > C p.S364L c.1091C > T p.S365L c.1094C > T p.P367R c.1100C > G p.S394* c.1181C > G p.T401I c.1202C > T p.P403fs*8 c.1208delC p.A404fs*9 c.1208_1209insC p.G421V c.1262G > T p.G421G c.1263A > G p.K439Q c.1315A > C p.K439T c.1316A > C p.T440P c.1318A > C p.T440A c.1318A > G p.T440T c.1320A > G p.G442S c.1324G > A p.R444W c.1330C > T p.R444Q c.1331G > A p.R444L c.1331G > T p.R444R c.1332G > A p.R444R c.1332G > T p.S447S c.1341T > C p.W450* c.1349G > A p.W450L c.1349G > T p.P453T c.1357C > A p.P453P c.1359T > C p.G455R c.1363G > A p.G455E c.1364G > A p.Q456* c.1366C > T p.Q456R c.1367A > G p.Q456Q c.1368G > A p.I457T c.1370T > C p.V459L c.1375G > C p.V459A c.1376T > C p.V459V c.1377G > A p.G460* c.1378G > T p.G460G c.1380A > G p.R462G c.1384A > G p.R462K c.1385G > A p.R462I c.1385G > T p.R462R c.1386A > G p.I463V c.1387A > G p.I463S c.1388T > G p.I463I c.1389T > C p.G464R c.1390G > A p.G464R c.1390G > C p.G464E c.1391G > A p.G464V c.1391G > T p.S465S c.1395T > C p.G466R c.1396G > A p.G466R c.1396G > C p.G466E c.1397G > A p.G466A c.1397G > C p.G466V c.1397G > T p.G466G c.1398A > G p.S467P c.1399T > C p.S467L c.1400C > T p.F468L c.1402T > C p.F468S c.1403T > C p.F468C c.1403T > G p.F468F c.1404T > C p.G469R c.1405G > A p.G469R c.1405G > C p.G469>? c.1405_1406GG > CT p.G469S c.1405_1406GG > TC p.G469L c.1405_1406GG > TT p.G469S c.1405_1407GGA > AGC p.G469S c.1405_1407GGA > AGT p.G469E c.1406G > A p.G469A c.1406G > C p.G469V c.1406G > T p.G469G c.1407A > G p.V471I c.1411G > A p.V471F c.1411G > T p.V471A c.1412T > C p.Y472S c.1415A > C p.Y472C c.1415A > G p.K475R c.1424A > G p.K475M c.1424A > T p.K475K c.1425G > A p.D479Y c.1435G > T p.L485L c.1453T > C p.L485S c.1454T > C p.L485_P490 > Y c.1454_1469 > A p.L485F c.1455G > T p.N486_P490del c.1457_1471del15 p.V487V c.1461G > A p.L505H c.1514T > A p.R509* c.1525C > T p.L514P c.1541T > C p.W531C c.1593G > T p.L537S c.1610T > C p.H539P c.1616A > C p.H542Y c.1624C > T p.K570K c.1710G > A p.H574N c.1720C > A p.H574Q c.1722C > A p.N581S c.1742A > G p.N581I c.1742A > T p.I582M c.1746A > G p.F583S c.1748T > C p.F583F c.1749T > C p.L584F c.1750C > T p.L584P c.1751T > C p.L584L c.1752T > C p.H585H c.1755T > C p.E586K c.1756G > A p.E586E c.1758A > G p.D587N c.1759G > A p.D587A c.1760A > C p.D587G c.1760A > G p.D587E c.1761C > A p.D587E c.1761C > G p.L588P c.1763T > C p.L588R c.1763T > G p.L588L c.1764C > T p.T589A c.1765A > G p.T589I c.1766C > T p.T589T c.1767A > G p.V590I c.1768G > A p.V590A c.1769T > C p.V590fs*3 c.1769delT p.V590V c.1770A > G p.K591E c.1771A > G p.K591R c.1772A > G p.I592V c.1774A > G p.I592M c.1776A > G p.I592I c.1776A > T p.G593S c.1777G > A p.G593C c.1777G > T p.G593D c.1778G > A p.D594N c.1779_1780TG > GA p.D594N c.1780G > A p.D594H c.1780G > C p.D594G c.1781A > G p.D594V c.1781A > T p.D594E c.1782T > A p.D594D c.1782T > C p.D594E c.1782T > G p.F595L c.1783T > C p.F595S c.1784T > C p.F595L c.1785T > A p.F595F c.1785T > C p.F595L c.1785T > G p.G596R c.1786G > C p.G596fs*2 c.1786delG p.G596D c.1787G > A p.G596G c.1788T > C p.L597V c.1789C > G p.L597S c.1789_1790CT > TC p.L597Q c.1790T > A p.L597P c.1790T > C p.L597R c.1790T > G p.L597L c.1791A > G p.A598T c.1792G > A p.A598V c.1793C > T p.A598A c.1794T > A p.A598_T599insV c.1794_1795insGTT p.T599del c.1794_1796delTAC p.T599I c.1796C > T p.T599_V600insT c.1796_1797insTAC p.T599_V600 > IAL c.1796_1798CAG > TAGCTT p.T599_R603 > I c.1796 1809 > TC p.T599T c.1797A > B p.T599T c.1797A > G p.T599T c.1797A > T p.T599_V600insTT c.1797_1797A > TACTACG p.T599_V600insTT c.1797_1798ins? p.T599_V600insT c.1797_1798insACA p.T599_V600insDFGLAT c.1798_1799ins18 p.V600R c.1797_1799AGT > GAG p.V600M c.1798G > A p.V600L c.1798G > C p.V600L c.1798G > T p.V600 > YM c.1798_1798G > TACA p.V600K c.1798_1799GT > AA p.V600R c.1798_1799GT > AG p.V600Q c.1798_1799GT > CA p.V600E c.1799T > A p.V600A c.1799T > C p.V600G c.1799T > G p.V600E c.1799_1800TG > AA p.V600D c.1799_1800TG > AC p.V600D c.1799_1800TG > AT p.V600fs*11 c.1799_1800delTG p.V600_K601 > E c.1799_1801delTGA p.V600_S602 > DT c.1799_1804TGAAAT > ATA p.V600_S605 > D c.1799_1814 > A p.V600_S605 > DV c.1799_1814 > ATGT p.V600_S605 > EK c.1799_1815 > AAAAG p.V600V c.1800G > A p.V600? c.(1798-1800)? p.K601E c.1801A > G p.K601del c.1801_1803delAAA p.K601R c.1802A > G p.K601I c.1802A > T p.K601N c.1803A > C p.K601K c.1803A > G p.K601N c.1803A > T p.S602S c.1806T > G p.R603R c.1807C > A p.R603* c.1807C > T p.R603L c.1808G > T p.R603R c.1809A > G p.W604del c.1808_1810delGAT p.W604R c.1810T > A p.W604G c.1810T > G p.W604* c.1811G > A p.W604* c.1812G > A p.S605G c.1813A > G p.S605F c.1813_1814AG > TT p.S605N c.1814G > A p.S605R c.1815T > A p.G606R c.1816G > A p.G606S c.1816_1818GGG > AGT p.G606E c.1817G > A p.G606A c.1817G > C p.G606V c.1817G > T p.G606G c.1818G > A p.S607P c.1819T > C p.H608R c.1823A > G p.H608H c.1824T > C p.Q609R c.1826A > G p.Q609Q c.1827G > A p.F610L c.1828T > C p.F610S c.1829T > C p.F610F c.1830T > C p.E611G c.1832A > G p.E611D c.1833A > C p.E611E c.1833A > G p.Q612E c.1834C > G p.Q612* c.1834C > T p.S614P c.1840T > C p.S614S c.1842T > C p.G615R c.1843G > A p.S616P c.1846T > C p.S616F c.1847C > T p.I617T c.1850T > C p.L618L c.1852T > C p.L618S c.1853T > C p.L618W c.1853T > G p.W619R c.1855T > C p.Q636E c.1906C > G p.Q636* c.1906C > T p.Q636R c.1907A > G p.S637P c.1909T > C p.S637* c.1910C > G p.S637L c.1910C > T p.S657S c.1971A > G p.R671Q c.2012G > A p.R682W c.2044C > T p.R682Q c.2045G > A p.K698R c.2093A > G p.A718V c.2153C > T p.P731S c.2191C > T p.P731P c.2193C > T

TABLE 7 ALK Capture Primers List for NGS Panel - Genomic DNA Seq. ID Primer Sequence ALK Exon21 130-150 bases 1 Left CCTCTTGTCTTCTCCTTTGCAC 2 Right GGGCAGGCTCAAGAGTGA 3 Left CCTCTTGTCTTCTCCTTTGCAC 4 Right AGGGCAGGCTCAAGAGTGA 5 Left CCTCTTGTCTTCTCCTTTGCAC 6 Right AAGGGCAGGCTCAAGAGTGA 7 Left CCTCTTGTCTTCTCCTTTGCAC 8 Right CAAGGGCAGGCTCAAGAGTGA 9 Left CTCTTGTCTTCTCCTTTGCAC 10 Right CAAGGGCAGGCTCAAGAGT 11 Left CTCTTGTCTTCTCCTTTGCAC 12 Right AAGGGCAGGCTCAAGAGTG 13 Left CTCTTGTCTTCTCCTTTGCAC 14 Right GGGCAGGCTCAAGAGTGA 15 Left CTCTTGTCTTCTCCTTTGCAC 16 Right AGGGCAGGCTCAAGAGTGA 17 Left CCTCTTGTCTTCTCCTTTGCAC 18 Right CCAAGGGCAGGCTCAAGAGTGA 19 Left CTCTTGTCTTCTCCTTTGCAC 20 Right AAGGGCAGGCTCAAGAGTGA 21 Left CCTCTTGTCTTCTCCTTTGCAC 22 Right AGCCAAGGGCAGGCTCAA 23 Left CTCTTGTCTTCTCCTTTGCAC 24 Right AGGGCAGGCTCAAGAGTG 25 Left CCTCTTGTCTTCTCCTTTGC 26 Right GGGCAGGCTCAAGAGTGA 27 Left CCTCTTGTCTTCTCCTTTGC 28 Right AGGGCAGGCTCAAGAGTGA 29 Left CTCTTGTCTTCTCCTTTGCAC 30 Right CAAGGGCAGGCTCAAGAGTG 31 Left CCTCTTGTCTTCTCCTTTGC 32 Right AAGGGCAGGCTCAAGAGTGA 33 Left CCTCTTGTCTTCTCCTTTGCAC 34 Right AGCCAAGGGCAGGCTCAAGAGTGA 35 Left TCTTGTCTTCTCCTTTGCAC 36 Right CAAGGGCAGGCTCAAGAGT 37 Left TCTTGTCTTCTCCTTTGCAC 38 Right AAGGGCAGGCTCAAGAGTG 39 Left CTCTTGTCTTCTCCTTTGCAC 40 Right CCAAGGGCAGGCTCAAGAGT ALK Exon21 151-200 bases 41 Left ACTCTGTCTCCTCTTGTCTTCTCCT 42 Right CTGAGAACTGCAGCCTACAGAGT 43 Left CTCTGTCTCCTCTTGTCTTCTCCTT 44 Right CTGAGAACTGCAGCCTACAGAGT 45 Left TCTGTCTCCTCTTGTCTTCTCCTT 46 Right CTGAGAACTGCAGCCTACAGAGT 47 Left TCTGTCTCCTCTTGTCTTCTCCTTT 48 Right CTGAGAACTGCAGCCTACAGAGT 49 Left ACTCTGTCTCCTCTTGTCTTCTCCT 50 Right AGAACTGCAGCCTACAGAGTCC 51 Left CTCTGTCTCCTCTTGTCTTCTCCT 52 Right CTGAGAACTGCAGCCTACAGAGT 53 Left TTGACTCTGTCTCCTCTTGTCTTCT 54 Right CTGAGAACTGCAGCCTACAGAG 55 Left CTGTCTCCTCTTGTCTTCTCCTTT 56 Right CTGAGAACTGCAGCCTACAGAGT 57 Left CTCTGTCTCCTCTTGTCTTCTCCTT 58 Right AGAACTGCAGCCTACAGAGTCC 59 Left ACTCTGTCTCCTCTTGTCTTCTCCT 60 Right CTGAGAACTGCAGCCTACAGAG 61 Left GTTTGACTCTGTCTCCTCTTGTCTT 62 Right CTGAGAACTGCAGCCTACAGAG 63 Left TCTGTCTCCTCTTGTCTTCTCCTT 64 Right AGAACTGCAGCCTACAGAGTCC 65 Left ACTCTGTCTCCTCTTGTCTTCTCCT 66 Right GAGAACTGCAGCCTACAGAGTCC 67 Left TCTGTCTCCTCTTGTCTTCTCCTTT 68 Right AGAACTGCAGCCTACAGAGTCC 69 Left CTGTCTCCTCTTGTCTTCTCCTTTG 70 Right CTGAGAACTGCAGCCTACAGAGT 71 Left CTCTGTCTCCTCTTGTCTTCTCCT 72 Right AGAACTGCAGCCTACAGAGTCC 73 Left CTGTTTGACTCTGTCTCCTCTTGTC 74 Right CTGAGAACTGCAGCCTACAGAG 75 Left CTCTGTCTCCTCTTGTCTTCTCCTT 76 Right CTGAGAACTGCAGCCTACAGAG 77 Left CTGTCTCCTCTTGTCTTCTCCTTT 78 Right AGAACTGCAGCCTACAGAGTCC 79 Left ACTCTGTCTCCTCTTGTCTTCTCC 80 Right AGAACTGCAGCCTACAGAGTCC ALK Exon21 201-300 bases 81 Left TGTTGAGGGTATTACTCCTGAGTGT 82 Right CTGAGAACTGCAGCCTACAGAGT 83 Left TTGAGGGTATTACTCCTGAGTGTGT 84 Right CTGAGAACTGCAGCCTACAGAGT 85 Left GTTGAGGGTATTACTCCTGAGTGTG 86 Right AGAACTGCAGCCTACAGAGTCC 87 Left TGTTGAGGGTATTACTCCTGAGTGT 88 Right AGAACTGCAGCCTACAGAGTCC 89 Left TTGAGGGTATTACTCCTGAGTGTGT 90 Right AGAACTGCAGCCTACAGAGTCC 91 Left CTCTCGTGTTTGTCCACTAAATGT 92 Right CTGAGAACTGCAGCCTACAGAGT 93 Left TGAGGGTATTACTCCTGAGTGTGTAT 94 Right CTGAGAACTGCAGCCTACAGAGT 95 Left GTTGAGGGTATTACTCCTGAGTGTG 96 Right CTGAGAACTGCAGCCTACAGAG 97 Left TGTTGAGGGTATTACTCCTGAGTGT 98 Right CTGAGAACTGCAGCCTACAGAG 99 Left TTGAGGGTATTACTCCTGAGTGTGT 100 Right CTGAGAACTGCAGCCTACAGAG 101 Left TGTTGAGGGTATTACTCCTGAGTGT 102 Right TGAGAACTGCAGCCTACAGAGT 103 Left TTGAGGGTATTACTCCTGAGTGTGT 104 Right TGAGAACTGCAGCCTACAGAGT 105 Left TGAGGGTATTACTCCTGAGTGTGT 106 Right CTGAGAACTGCAGCCTACAGAGT 107 Left GTTGAGGGTATTACTCCTGAGTGTG 108 Right GAGAACTGCAGCCTACAGAGTCC 109 Left TGTTGAGGGTATTACTCCTGAGTGT 110 Right GAGAACTGCAGCCTACAGAGTCC 111 Left TTGAGGGTATTACTCCTGAGTGTGT 112 Right GAGAACTGCAGCCTACAGAGTCC 113 Left GTTGAGGGTATTACTCCTGAGTGTGT 114 Right CTGAGAACTGCAGCCTACAGAGT 115 Left CTCTCGTGTTTGTCCACTAAATGTG 116 Right CTGAGAACTGCAGCCTACAGAGT 117 Left TTGACTCTGTCTCCTCTTGTCTTCT 118 Right GAGGCTGTGAGCTGAGAACTG 119 Left CTCTCGTGTTTGTCCACTAAATGT 120 Right AGAACTGCAGCCTACAGAGTCC ALK Exon21 301-400 bases 121 Left GAATCCTTCTTACCAGTTTTCAGGT 122 Right CTGAGAACTGCAGCCTACAGAGT 123 Left GTTGGAATCCTTCTTACCAGTTTTC 124 Right CTGAGAACTGCAGCCTACAGAGT 125 Left AATCCTTCTTACCAGTTTTCAGGTG 126 Right CTGAGAACTGCAGCCTACAGAGT 127 Left ATCCTTCTTACCAGTTTTCAGGTG 128 Right CTGAGAACTGCAGCCTACAGAGT 129 Left GAATCCTTCTTACCAGTTTTCAGGT 130 Right AGAACTGCAGCCTACAGAGTCC 131 Left TTGGAATCCTTCTTACCAGTTTTC 132 Right CTGAGAACTGCAGCCTACAGAGT 133 Left GTTGGAATCCTTCTTACCAGTTTTC 134 Right AGAACTGCAGCCTACAGAGTCC 135 Left GAATCCTTCTTACCAGTTTTCAGG 136 Right CTGAGAACTGCAGCCTACAGAGT 137 Left GGAATCCTTCTTACCAGTTTTCAG 138 Right CTGAGAACTGCAGCCTACAGAGT 139 Left ATGTTGAGGGTATTACTCCTGAGTGT 140 Right CTGAGAACTGCAGCCTACAGAGT 141 Left GAATCCTTCTTACCAGTTTTCAGGT 142 Right CTGAGAACTGCAGCCTACAGAG 143 Left CAAAGCCATGTTGAGGGTATTACT 144 Right CTGAGAACTGCAGCCTACAGAGT 145 Left GAATCCTTCTTACCAGTTTTCAGGT 146 Right TGAGAACTGCAGCCTACAGAGT 147 Left GAATCCTTCTTACCAGTTTTCAGGT 148 Right GAGAACTGCAGCCTACAGAGTCC 149 Left GTTGGAATCCTTCTTACCAGTTTTC 150 Right CTGAGAACTGCAGCCTACAGAG 151 Left AATCCTTCTTACCAGTTTTCAGGTG 152 Right AGAACTGCAGCCTACAGAGTCC 153 Left ATCCTTCTTACCAGTTTTCAGGTG 154 Right AGAACTGCAGCCTACAGAGTCC 155 Left GGTTGGAATCCTTCTTACCAGTTT 156 Right CTGAGAACTGCAGCCTACAGAGT 157 Left TGGAATCCTTCTTACCAGTTTTCAG 158 Right CTGAGAACTGCAGCCTACAGAGT 159 Left TTGGAATCCTTCTTACCAGTTTTC 160 Right AGAACTGCAGCCTACAGAGTCC ALK Exon21-22 301-400 bases 161 Left TTGACTCTGTCTCCTCTTGTCTTCT 162 Right TGGAGATATCGATCTGTTAGAAACC 163 Left TTTGACTCTGTCTCCTCTTGTCTTC 164 Right CCTTGGAGATATCGATCTGTTAGAA 165 Left ACTCTGTCTCCTCTTGTCTTCTCCT 166 Right CCTTGGAGATATCGATCTGTTAGAA 167 Left GTTTGACTCTGTCTCCTCTTGTCTT 168 Right CCTTGGAGATATCGATCTGTTAGAA 169 Left TTTGACTCTGTCTCCTCTTGTCTTC 170 Right TGGAGATATCGATCTGTTAGAAACC 171 Left ACTCTGTCTCCTCTTGTCTTCTCCT 172 Right TGGAGATATCGATCTGTTAGAAACC 173 Left TTGACTCTGTCTCCTCTTGTCTTCT 174 Right TATCGATCTGTTAGAAACCTCTCCA 175 Left TGACTCTGTCTCCTCTTGTCTTCTC 176 Right CCTTGGAGATATCGATCTGTTAGAA 177 Left GTTTGACTCTGTCTCCTCTTGTCTT 178 Right TGGAGATATCGATCTGTTAGAAACC 179 Left TGACTCTGTCTCCTCTTGTCTTCTC 180 Right TGGAGATATCGATCTGTTAGAAACC 181 Left TGTTTGACTCTGTCTCCTCTTGTCT 182 Right CCTTGGAGATATCGATCTGTTAGAA 183 Left CTGTTTGACTCTGTCTCCTCTTGTC 184 Right CCTTGGAGATATCGATCTGTTAGAA 185 Left CTCTGTCTCCTCTTGTCTTCTCCTT 186 Right CCTTGGAGATATCGATCTGTTAGAA 187 Left TTTGACTCTGTCTCCTCTTGTCTTC 188 Right TATCGATCTGTTAGAAACCTCTCCA 189 Left ACTCTGTCTCCTCTTGTCTTCTCCT 190 Right TATCGATCTGTTAGAAACCTCTCCA 191 Left TTGACTCTGTCTCCTCTTGTCTTCT 192 Right GTTAGAAACCTCTCCAGGTTCTTTG 193 Left GTTTGACTCTGTCTCCTCTTGTCTT 194 Right TATCGATCTGTTAGAAACCTCTCCA 195 Left TGTTTGACTCTGTCTCCTCTTGTCT 196 Right TGGAGATATCGATCTGTTAGAAACC 197 Left CTGTTTGACTCTGTCTCCTCTTGTC 198 Right TGGAGATATCGATCTGTTAGAAACC 199 Left CTCTGTCTCCTCTTGTCTTCTCCTT 200 Right TGGAGATATCGATCTGTTAGAAACC ALK Exon21-22 401-500 bases 201 Left TTGACTCTGTCTCCTCTTGTCTTCT 202 Right TAGAATGTTTGGGAGTCTCCTACTG 203 Left TTGACTCTGTCTCCTCTTGTCTTCT 204 Right GTTGTTCCATTCTGGTAAGAAGTGT 205 Left GTTGAGGGTATTACTCCTGAGTGTG 206 Right CCTTGGAGATATCGATCTGTTAGAA 207 Left TGTTGAGGGTATTACTCCTGAGTGT 208 Right CCTTGGAGATATCGATCTGTTAGAA 209 Left TTGAGGGTATTACTCCTGAGTGTGT 210 Right CCTTGGAGATATCGATCTGTTAGAA 211 Left GTTGAGGGTATTACTCCTGAGTGTG 212 Right TGGAGATATCGATCTGTTAGAAACC 213 Left TGTTGAGGGTATTACTCCTGAGTGT 214 Right TGGAGATATCGATCTGTTAGAAACC 215 Left TTGAGGGTATTACTCCTGAGTGTGT 216 Right TGGAGATATCGATCTGTTAGAAACC 217 Left TTGACTCTGTCTCCTCTTGTCTTCT 218 Right GAAGTGTCTAGAATGTTTGGGAGTC 219 Left TTTGACTCTGTCTCCTCTTGTCTTC 220 Right TAGAATGTTTGGGAGTCTCCTACTG 221 Left ACTCTGTCTCCTCTTGTCTTCTCCT 222 Right TAGAATGTTTGGGAGTCTCCTACTG 223 Left TTGACTCTGTCTCCTCTTGTCTTCT 224 Right TGTTCCATTCTGGTAAGAAGTGTCT 225 Left GTTTGACTCTGTCTCCTCTTGTCTT 226 Right TAGAATGTTTGGGAGTCTCCTACTG 227 Left TTTGACTCTGTCTCCTCTTGTCTTC 228 Right GTTGTTCCATTCTGGTAAGAAGTGT 229 Left ACTCTGTCTCCTCTTGTCTTCTCCT 230 Right GTTGTTCCATTCTGGTAAGAAGTGT 231 Left GTTTGACTCTGTCTCCTCTTGTCTT 232 Right GTTGTTCCATTCTGGTAAGAAGTGT 233 Left TGACTCTGTCTCCTCTTGTCTTCTC 234 Right TAGAATGTTTGGGAGTCTCCTACTG 235 Left TGACTCTGTCTCCTCTTGTCTTCTC 236 Right GTTGTTCCATTCTGGTAAGAAGTGT 237 Left GTTGAGGGTATTACTCCTGAGTGTG 238 Right TATCGATCTGTTAGAAACCTCTCCA 239 Left TGTTGAGGGTATTACTCCTGAGTGT 240 Right TATCGATCTGTTAGAAACCTCTCCA ALK Exon21-22 501-600 bases 241 Left GAATCCTTCTTACCAGTTTTCAGGT 242 Right CCTTGGAGATATCGATCTGTTAGAA 243 Left GAATCCTTCTTACCAGTTTTCAGGT 244 Right TGGAGATATCGATCTGTTAGAAACC 245 Left GTTGAGGGTATTACTCCTGAGTGTG 246 Right TAGAATGTTTGGGAGTCTCCTACTG 247 Left TGTTGAGGGTATTACTCCTGAGTGT 248 Right TAGAATGTTTGGGAGTCTCCTACTG 249 Left TTGAGGGTATTACTCCTGAGTGTGT 250 Right TAGAATGTTTGGGAGTCTCCTACTG 251 Left GTTGAGGGTATTACTCCTGAGTGTG 252 Right GTTGTTCCATTCTGGTAAGAAGTGT 253 Left TGTTGAGGGTATTACTCCTGAGTGT 254 Right GTTGTTCCATTCTGGTAAGAAGTGT 255 Left TTGAGGGTATTACTCCTGAGTGTGT 256 Right GTTGTTCCATTCTGGTAAGAAGTGT 257 Left GTTGGAATCCTTCTTACCAGTTTTC 258 Right CCTTGGAGATATCGATCTGTTAGAA 259 Left GTTGAGGGTATTACTCCTGAGTGTG 260 Right GAAGTGTCTAGAATGTTTGGGAGTC 261 Left TTGAGGGTATTACTCCTGAGTGTGT 262 Right GAAGTGTCTAGAATGTTTGGGAGTC 263 Left TGTTGAGGGTATTACTCCTGAGTGT 264 Right GAAGTGTCTAGAATGTTTGGGAGTC 265 Left GAATCCTTCTTACCAGTTTTCAGGT 266 Right TATCGATCTGTTAGAAACCTCTCCA 267 Left GTTGAGGGTATTACTCCTGAGTGTG 268 Right TGTTCCATTCTGGTAAGAAGTGTCT 269 Left TGTTGAGGGTATTACTCCTGAGTGT 270 Right TGTTCCATTCTGGTAAGAAGTGTCT 271 Left TTGAGGGTATTACTCCTGAGTGTGT 272 Right TGTTCCATTCTGGTAAGAAGTGTCT 273 Left AATCCTTCTTACCAGTTTTCAGGTG 274 Right CCTTGGAGATATCGATCTGTTAGAA 275 Left GTTGGAATCCTTCTTACCAGTTTTC 276 Right TATCGATCTGTTAGAAACCTCTCCA 277 Left ATCCTTCTTACCAGTTTTCAGGTG 278 Right CCTTGGAGATATCGATCTGTTAGAA 279 Left GTTGAGGGTATTACTCCTGAGTGTG 280 Right TTGTTCCATTCTGGTAAGAAGTGTC ALK Exon21-22 601-800 bases 281 Left TTGACTCTGTCTCCTCTTGTCTTCT 282 Right AAAGTCTAGCATGCTCCATTTCTTA 283 Left GAATCCTTCTTACCAGTTTTCAGGT 284 Right TAGAATGTTTGGGAGTCTCCTACTG 285 Left GAATCCTTCTTACCAGTTTTCAGGT 286 Right AAAGTCTAGCATGCTCCATTTCTTA 287 Left GTTGAGGGTATTACTCCTGAGTGTG 288 Right AAAGTCTAGCATGCTCCATTTCTTA 289 Left TGTTGAGGGTATTACTCCTGAGTGT 290 Right AAAGTCTAGCATGCTCCATTTCTTA 291 Left TTGAGGGTATTACTCCTGAGTGTGT 292 Right AAAGTCTAGCATGCTCCATTTCTTA 293 Left CCTCTGTCACTCACTGGAAATACTC 294 Right CCTTGGAGATATCGATCTGTTAGAA 295 Left TCCTCTGTCACTCACTGGAAATACT 296 Right CCTTGGAGATATCGATCTGTTAGAA 297 Left CTCTGTCACTCACTGGAAATACTCC 298 Right CCTTGGAGATATCGATCTGTTAGAA 299 Left TTTGACTCTGTCTCCTCTTGTCTTC 300 Right AAAGTCTAGCATGCTCCATTTCTTA 301 Left ACTCTGTCTCCTCTTGTCTTCTCCT 302 Right AAAGTCTAGCATGCTCCATTTCTTA 303 Left TTGACTCTGTCTCCTCTTGTCTTCT 304 Right GGTCTTGGAGGGAGATTATATCTTG 305 Left GTTGGAATCCTTCTTACCAGTTTTC 306 Right TAGAATGTTTGGGAGTCTCCTACTG 307 Left GTTTGACTCTGTCTCCTCTTGTCTT 308 Right AAAGTCTAGCATGCTCCATTTCTTA 309 Left GAATCCTTCTTACCAGTTTTCAGGT 310 Right GAAGTGTCTAGAATGTTTGGGAGTC 311 Left CCTCTGTCACTCACTGGAAATACTC 312 Right TGGAGATATCGATCTGTTAGAAACC 313 Left TCCTCTGTCACTCACTGGAAATACT 314 Right TGGAGATATCGATCTGTTAGAAACC 315 Left CTCTGTCACTCACTGGAAATACTCC 316 Right TGGAGATATCGATCTGTTAGAAACC 317 Left GTTGGAATCCTTCTTACCAGTTTTC 318 Right AAAGTCTAGCATGCTCCATTTCTTA 319 Left TGACTCTGTCTCCTCTTGTCTTCTC 320 Right AAAGTCTAGCATGCTCCATTTCTTA ALK Exon21-22 801-1000 bases 321 Left CTCTCCTCAAAATTCATTCAGATGT 322 Right CCTTGGAGATATCGATCTGTTAGAA 323 Left ATGTTGGCTTACATTAACTCCCATA 324 Right CCTTGGAGATATCGATCTGTTAGAA 325 Left CTCTCCTCAAAATTCATTCAGATGT 326 Right TAGAATGTTTGGGAGTCTCCTACTG 327 Left ATGTTGGCTTACATTAACTCCCATA 328 Right TAGAATGTTTGGGAGTCTCCTACTG 329 Left CTCTCCTCAAAATTCATTCAGATGT 330 Right TGGAGATATCGATCTGTTAGAAACC 331 Left CTCTCCTCAAAATTCATTCAGATGT 332 Right GTTGTTCCATTCTGGTAAGAAGTGT 333 Left ATGTTGGCTTACATTAACTCCCATA 334 Right TGGAGATATCGATCTGTTAGAAACC 335 Left AAAATTCATTCAGATGTGCTCTCTC 336 Right TAGAATGTTTGGGAGTCTCCTACTG 337 Left AAAATTCATTCAGATGTGCTCTCTC 338 Right TGGAGATATCGATCTGTTAGAAACC 339 Left AAAATTCATTCAGATGTGCTCTCTC 340 Right GTTGTTCCATTCTGGTAAGAAGTGT 341 Left CTCTCCTCAAAATTCATTCAGATGT 342 Right GAAGTGTCTAGAATGTTTGGGAGTC 343 Left CTCTCCTCAAAATTCATTCAGATGT 344 Right TGTTCCATTCTGGTAAGAAGTGTCT 345 Left CTCTCCTCAAAATTCATTCAGATGT 346 Right TATCGATCTGTTAGAAACCTCTCCA 347 Left ATGTTGGCTTACATTAACTCCCATA 348 Right TATCGATCTGTTAGAAACCTCTCCA 349 Left AAAATTCATTCAGATGTGCTCTCTC 350 Right GAAGTGTCTAGAATGTTTGGGAGTC 351 Left AAAATTCATTCAGATGTGCTCTCTC 352 Right TGTTCCATTCTGGTAAGAAGTGTCT 353 Left TGGCTTACATTAACTCCCATAGTTT 354 Right CCTTGGAGATATCGATCTGTTAGAA 355 Left TTGGCTTACATTAACTCCCATAGTT 356 Right CCTTGGAGATATCGATCTGTTAGAA 357 Left TGTTGGCTTACATTAACTCCCATAG 358 Right CCTTGGAGATATCGATCTGTTAGAA 359 Left AAAATTCATTCAGATGTGCTCTCTC 360 Right TATCGATCTGTTAGAAACCTCTCCA ALK Exon21-22 2 kb 361 Left CTCTCCTCAAAATTCATTCAGATGT 362 Right GCAGGAGAGTGTCTTTCTCAGATAC 363 Left ATGTTGGCTTACATTAACTCCCATA 364 Right GCAGGAGAGTGTCTTTCTCAGATAC 365 Left AAAATTCATTCAGATGTGCTCTCTC 366 Right GCAGGAGAGTGTCTTTCTCAGATAC 367 Left CTCTCCTCAAAATTCATTCAGATGT 368 Right GGAGAGTGTCTTTCTCAGATACTGG 369 Left ATGTTGGCTTACATTAACTCCCATA 370 Right GGAGAGTGTCTTTCTCAGATACTGG 371 Left AAAATTCATTCAGATGTGCTCTCTC 372 Right CAAAGTTACATTTTCAGCAGCTACA 373 Left AAAATTCATTCAGATGTGCTCTCTC 374 Right GGAGAGTGTCTTTCTCAGATACTGG 375 Left GAATCCTTCTTACCAGTTTTCAGGT 376 Right CAAAGTTACATTTTCAGCAGCTACA 377 Left CTCTCCTCAAAATTCATTCAGATGT 378 Right GCAGCTACAATGTATAAAGGCATTC 379 Left GTTGAGGGTATTACTCCTGAGTGTG 380 Right CAAAGTTACATTTTCAGCAGCTACA 381 Left TGTTGAGGGTATTACTCCTGAGTGT 382 Right CAAAGTTACATTTTCAGCAGCTACA 383 Left ATGTTGGCTTACATTAACTCCCATA 384 Right TTAACATGATCCCTTTAGGACACAC 385 Left ATGTTGGCTTACATTAACTCCCATA 386 Right TGTTAACATGATCCCTTTAGGACAC 387 Left ATGTTGGCTTACATTAACTCCCATA 388 Right GTTAACATGATCCCTTTAGGACACA 389 Left AAAATTCATTCAGATGTGCTCTCTC 390 Right GCAGCTACAATGTATAAAGGCATTC 391 Left CTCTGTCACTCACTGGAAATACTCC 392 Right GCAGGAGAGTGTCTTTCTCAGATAC 393 Left TCCTCTGTCACTCACTGGAAATACT 394 Right GCAGGAGAGTGTCTTTCTCAGATAC 395 Left TGGCTTACATTAACTCCCATAGTTT 396 Right GCAGGAGAGTGTCTTTCTCAGATAC 397 Left TTGGCTTACATTAACTCCCATAGTT 398 Right GCAGGAGAGTGTCTTTCTCAGATAC 399 Left TGTTGGCTTACATTAACTCCCATAG 400 Right GCAGGAGAGTGTCTTTCTCAGATAC ALK Exon22 90-150 bases 401 Left AGTTCTCAGCTCACAGCCTCCT 402 Right AGGGTGTCTCTCTGTGGCTTTAC 403 Left AGTTCTCAGCTCACAGCCTCCT 404 Right GGGTGTCTCTCTGTGGCTTTAC 405 Left GTTCTCAGCTCACAGCCTCCT 406 Right AGGGTGTCTCTCTGTGGCTTTAC 407 Left TAGGCTGCAGTTCTCAGCTCAC 408 Right AGGGTGTCTCTCTGTGGCTTTAC 409 Left GTTCTCAGCTCACAGCCTCCT 410 Right GGGTGTCTCTCTGTGGCTTTAC 411 Left AGTTCTCAGCTCACAGCCTCCT 412 Right AGGGTGTCTCTCTGTGGCTTTA 413 Left TAGGCTGCAGTTCTCAGCTCAC 414 Right GGGTGTCTCTCTGTGGCTTTAC 415 Left TTCTCAGCTCACAGCCTCCT 416 Right AGGGTGTCTCTCTGTGGCTTTAC 417 Left GTTCTCAGCTCACAGCCTCCT 418 Right AGGGTGTCTCTCTGTGGCTTTA 419 Left TTCTCAGCTCACAGCCTCCT 420 Right GGGTGTCTCTCTGTGGCTTTAC 421 Left TAGGCTGCAGTTCTCAGCTCAC 422 Right AGGGTGTCTCTCTGTGGCTTTA 423 Left GCTGCAGTTCTCAGCTCACAG 424 Right AGGGTGTCTCTCTGTGGCTTTAC 425 Left TTCTCAGCTCACAGCCTCCT 426 Right AGGGTGTCTCTCTGTGGCTTTA 427 Left GCTGCAGTTCTCAGCTCACAG 428 Right GGGTGTCTCTCTGTGGCTTTAC 429 Left AGTTCTCAGCTCACAGCCTCCT 430 Right GAGGGTGTCTCTCTGTGGCTTTAC 431 Left AGTTCTCAGCTCACAGCCTCCTC 432 Right AGGGTGTCTCTCTGTGGCTTTAC 433 Left GTTCTCAGCTCACAGCCTCCT 434 Right GAGGGTGTCTCTCTGTGGCTTTAC 435 Left TAGGCTGCAGTTCTCAGCTCAC 436 Right GAGGGTGTCTCTCTGTGGCTTTAC 437 Left AGTTCTCAGCTCACAGCCTCCTC 438 Right GGGTGTCTCTCTGTGGCTTTAC 439 Left TCTCAGCTCACAGCCTCCTC 440 Right AGGGTGTCTCTCTGTGGCTTTAC ALK Exon22 151-200 bases 441 Left AGTTCTCAGCTCACAGCCTCCT 442 Right TATCGATCTGTTAGAAACCTCTCCA 443 Left GTTCTCAGCTCACAGCCTCCT 444 Right TATCGATCTGTTAGAAACCTCTCCA 445 Left TTCTCAGCTCACAGCCTCCT 446 Right TATCGATCTGTTAGAAACCTCTCCA 447 Left AGTTCTCAGCTCACAGCCTCCT 448 Right GTTAGAAACCTCTCCAGGTTCTTTG 449 Left GTTCTCAGCTCACAGCCTCCT 450 Right GTTAGAAACCTCTCCAGGTTCTTTG 451 Left TCTCAGCTCACAGCCTCCTC 452 Right TGGAGATATCGATCTGTTAGAAACC 453 Left TAGGCTGCAGTTCTCAGCTCAC 454 Right GTTAGAAACCTCTCCAGGTTCTTTG 455 Left AGTTCTCAGCTCACAGCCTCCT 456 Right GATATCGATCTGTTAGAAACCTCTCC 457 Left TTCTCAGCTCACAGCCTCCT 458 Right GTTAGAAACCTCTCCAGGTTCTTTG 459 Left AGTTCTCAGCTCACAGCCTCCT 460 Right TTAGAAACCTCTCCAGGTTCTTTG 461 Left AGTTCTCAGCTCACAGCCTCCTC 462 Right TATCGATCTGTTAGAAACCTCTCCA 463 Left GTTCTCAGCTCACAGCCTCCT 464 Right GATATCGATCTGTTAGAAACCTCTCC 465 Left GTTCTCAGCTCACAGCCTCCT 466 Right TTAGAAACCTCTCCAGGTTCTTTG 467 Left AGTTCTCAGCTCACAGCCTCCT 468 Right ATCGATCTGTTAGAAACCTCTCCAG 469 Left TAGGCTGCAGTTCTCAGCTCAC 470 Right TTAGAAACCTCTCCAGGTTCTTTG 471 Left AGCTCACAGCCTCCTCCTC 472 Right CCTTGGAGATATCGATCTGTTAGAA 473 Left GCTGCAGTTCTCAGCTCACAG 474 Right GTTAGAAACCTCTCCAGGTTCTTTG 475 Left TCTCAGCTCACAGCCTCCTC 476 Right TATCGATCTGTTAGAAACCTCTCCA 477 Left TTCTCAGCTCACAGCCTCCT 478 Right GATATCGATCTGTTAGAAACCTCTCC 479 Left GTTCTCAGCTCACAGCCTCCTC 480 Right TATCGATCTGTTAGAAACCTCTCCA ALK Exon22 201-300 bases 481 Left GGACTCTGTAGGCTGCAGTTCTC 482 Right CCTTGGAGATATCGATCTGTTAGAA 483 Left GGACTCTGTAGGCTGCAGTTCTC 484 Right TAGAATGTTTGGGAGTCTCCTACTG 485 Left GGACTCTGTAGGCTGCAGTTCTC 486 Right TGGAGATATCGATCTGTTAGAAACC 487 Left GGACTCTGTAGGCTGCAGTTCTC 488 Right GAAGTGTCTAGAATGTTTGGGAGTC 489 Left GGACTCTGTAGGCTGCAGTTCTC 490 Right TATCGATCTGTTAGAAACCTCTCCA 491 Left GGACTCTGTAGGCTGCAGTTCTC 492 Right GTTAGAAACCTCTCCAGGTTCTTTG 493 Left GGACTCTGTAGGCTGCAGTTCTC 494 Right AAGAAGTGTCTAGAATGTTTGGGAGT 495 Left GGACTCTGTAGGCTGCAGTTCTC 496 Right TGGTAAGAAGTGTCTAGAATGTTTGG 497 Left GGACTCTGTAGGCTGCAGTTCTC 498 Right AGAATGTTTGGGAGTCTCCTACTG 499 Left GGACTCTGTAGGCTGCAGTTCTC 500 Right TCTAGAATGTTTGGGAGTCTCCTACT 501 Left GGACTCTGTAGGCTGCAGTTCTC 502 Right GATATCGATCTGTTAGAAACCTCTCC 503 Left GGACTCTGTAGGCTGCAGTTCTC 504 Right TTAGAAACCTCTCCAGGTTCTTTG 505 Left GGACTCTGTAGGCTGCAGTTCTC 506 Right AGAAGTGTCTAGAATGTTTGGGAGT 507 Left GGACTCTGTAGGCTGCAGTTCTC 508 Right AAGTGTCTAGAATGTTTGGGAGTCT 509 Left AGTTCTCAGCTCACAGCCTCCT 510 Right CCTTGGAGATATCGATCTGTTAGAA 511 Left AGTTCTCAGCTCACAGCCTCCT 512 Right TAGAATGTTTGGGAGTCTCCTACTG 513 Left AGTTCTCAGCTCACAGCCTCCT 514 Right TGGAGATATCGATCTGTTAGAAACC 515 Left GGACTCTGTAGGCTGCAGTTCTC 516 Right ATCGATCTGTTAGAAACCTCTCCAG 517 Left GTTCTCAGCTCACAGCCTCCT 518 Right CCTTGGAGATATCGATCTGTTAGAA 519 Left AGTTCTCAGCTCACAGCCTCCT 520 Right GTTGTTCCATTCTGGTAAGAAGTGT ALK Exon22 301-400 bases 521 Left GGACTCTGTAGGCTGCAGTTCTC 522 Right GTTGTTCCATTCTGGTAAGAAGTGT 523 Left GGACTCTGTAGGCTGCAGTTCTC 524 Right TGTTCCATTCTGGTAAGAAGTGTCT 525 Left GGACTCTGTAGGCTGCAGTTCTC 526 Right TTGTTCCATTCTGGTAAGAAGTGTC 527 Left GGACTCTGTAGGCTGCAGTTCTC 528 Right GGTTGTTCCATTCTGGTAAGAAGT 529 Left GGACTCTGTAGGCTGCAGTTCTC 530 Right GGATTATTAGGCCACACAGACTTT 531 Left GGACTCTGTAGGCTGCAGTTCTC 532 Right TGTTCCATTCTGGTAAGAAGTGTCTA 533 Left GGACTCTGTAGGCTGCAGTTCTC 534 Right TGTTCCATTCTGGTAAGAAGTGTC 535 Left GGACTCTGTAGGCTGCAGTTCTC 536 Right ATACTGGTTGCAGACAGTGACATC 537 Left GGACTCTGTAGGCTGCAGTTCTC 538 Right GATACTGGTTGCAGACAGTGACAT 539 Left GGACTCTGTAGGCTGCAGTTCTC 540 Right ATTAGGCCACACAGACTTTGTTTCT 541 Left GGACTCTGTAGGCTGCAGTTCTC 542 Right TTGTTCCATTCTGGTAAGAAGTGT 543 Left GGACTCTGTAGGCTGCAGTTCTC 544 Right GTTCCATTCTGGTAAGAAGTGTCTA 545 Left GGACTCTGTAGGCTGCAGTTCTC 546 Right GATACTGGTTGCAGACAGTGACATC 547 Left CGGACTCTGTAGGCTGCAGTT 548 Right GTTGTTCCATTCTGGTAAGAAGTGT 549 Left GGACTCTGTAGGCTGCAGTTCTC 550 Right TAGGCCACACAGACTTTGTTTCT 551 Left GGACTCTGTAGGCTGCAGTTCTC 552 Right TACTGGTTGCAGACAGTGACATC 553 Left GTAGGCTGCAGTTCTCAGCTCACAG 554 Right GTTGTTCCATTCTGGTAAGAAGTGT 555 Left AGTTCTCAGCTCACAGCCTCCT 556 Right GGATTATTAGGCCACACAGACTTT 557 Left GGACTCTGTAGGCTGCAGTTCTC 558 Right TTAGGCCACACAGACTTTGTTTCT 559 Left GGACTCTGTAGGCTGCAGTTCTC 560 Right ACAGTGACATCGGTGGGATTATTAG ALK Exon23 151-200 bases 561 Left TTAATTTTGGTTACATCCCTCTCTG 562 Right AGCAAAGACTGGTTCTCACTCAC 563 Left CAGACTCAGCTCAGTTAATTTTGGT 564 Right AGCAAAGACTGGTTCTCACTCAC 565 Left AGCTCAGTTAATTTTGGTTACATCC 566 Right AGCAAAGACTGGTTCTCACTCAC 567 Left AGACTCAGCTCAGTTAATTTTGGTT 568 Right AGCAAAGACTGGTTCTCACTCAC 569 Left CTCAGCTCAGTTAATTTTGGTTACA 570 Right AGCAAAGACTGGTTCTCACTCAC 571 Left TCAGCTCAGTTAATTTTGGTTACATC 572 Right AGCAAAGACTGGTTCTCACTCAC 573 Left CAGTTAATTTTGGTTACATCCCTCT 574 Right AGCAAAGACTGGTTCTCACTCAC 575 Left ACTCAGCTCAGTTAATTTTGGTTACA 576 Right AGCAAAGACTGGTTCTCACTCAC 577 Left CAGTTAATTTTGGTTACATCCCTCTC 578 Right AGCAAAGACTGGTTCTCACTCAC 579 Left TCAGTTAATTTTGGTTACATCCCTCT 580 Right AGCAAAGACTGGTTCTCACTCAC 581 Left GTTAATTTTGGTTACATCCCTCTCTG 582 Right AGCAAAGACTGGTTCTCACTCAC 583 Left CAGACTCAGCTCAGTTAATTTTGG 584 Right AGCAAAGACTGGTTCTCACTCAC 585 Left CAGCTCAGTTAATTTTGGTTACATC 586 Right AGCAAAGACTGGTTCTCACTCAC 587 Left TCAGCTCAGTTAATTTTGGTTACAT 588 Right AGCAAAGACTGGTTCTCACTCAC 589 Left TTAATTTTGGTTACATCCCTCTCTG 590 Right AACTGCAGCAAAGACTGGTTCT 591 Left TTAATTTTGGTTACATCCCTCTCTG 592 Right ACAACAACTGCAGCAAAGACTG 593 Left TTAATTTTGGTTACATCCCTCTCTG 594 Right CACAACAACTGCAGCAAAGACT 595 Left CTCAGCTCAGTTAATTTTGGTTACAT 596 Right AGCAAAGACTGGTTCTCACTCAC 597 Left TTAATTTTGGTTACATCCCTCTCTG 598 Right CAGCAAAGACTGGTTCTCACTCAC 599 Left AGTTAATTTTGGTTACATCCCTCTC 600 Right AGCAAAGACTGGTTCTCACTCAC ALK Exon23 201-300 bases 601 Left TGTAGCTGCTGAAAATGTAACTTTG 602 Right AGCAAAGACTGGTTCTCACTCAC 603 Left TTAATTTTGGTTACATCCCTCTCTG 604 Right CTGTCCAAGCCTAAAGTTGACAC 605 Left TATCCTGTTCCTCCCAGTTTAAGAT 606 Right AGCAAAGACTGGTTCTCACTCAC 607 Left ATGCCTTTATACATTGTAGCTGCTG 608 Right AGCAAAGACTGGTTCTCACTCAC 609 Left GCCTTTATACATTGTAGCTGCTGAA 610 Right AGCAAAGACTGGTTCTCACTCAC 611 Left GTATCCTGTTCCTCCCAGTTTAAGA 612 Right AGCAAAGACTGGTTCTCACTCAC 613 Left GCCTTTATACATTGTAGCTGCTGA 614 Right AGCAAAGACTGGTTCTCACTCAC 615 Left AGTTTAAGATTTGCCCAGACTCAG 616 Right AGCAAAGACTGGTTCTCACTCAC 617 Left CCCAGACTCAGCTCAGTTAATTTT 618 Right AGCAAAGACTGGTTCTCACTCAC 619 Left TTAATTTTGGTTACATCCCTCTCTG 620 Right GTCCAAGCCTAAAGTTGACACC 621 Left CAGTTAATTTTGGTTACATCCCTCT 622 Right CTGTCCAAGCCTAAAGTTGACAC 623 Left TTAATTTTGGTTACATCCCTCTCTG 624 Right CCTGTCCAAGCCTAAAGTTGAC 625 Left TATCCTGTTCCTCCCAGTTTAAGA 626 Right AGCAAAGACTGGTTCTCACTCAC 627 Left CTGTTCCTCCCAGTTTAAGATTTG 628 Right AGCAAAGACTGGTTCTCACTCAC 629 Left CAGAATGCCTTTATACATTGTAGCTG 630 Right AGCAAAGACTGGTTCTCACTCAC 631 Left CCCATGTTTACAGAATGCCTTTAT 632 Right AGCAAAGACTGGTTCTCACTCAC 633 Left GCTGCTGAAAATGTAACTTTGTATC 634 Right AGCAAAGACTGGTTCTCACTCAC 635 Left GTATCCTGTTCCTCCCAGTTTAAG 636 Right AGCAAAGACTGGTTCTCACTCAC 637 Left TGTAGCTGCTGAAAATGTAACTTTG 638 Right AACTGCAGCAAAGACTGGTTCT 639 Left ATCCTGTTCCTCCCAGTTTAAGAT 640 Right AGCAAAGACTGGTTCTCACTCAC 641 Left TTAATTTTGGTTACATCCCTCTCTG 642 Right TCAGCCATCATCTACCTCTATCTTC 643 Left CAGACTCAGCTCAGTTAATTTTGGT 644 Right TCAGCCATCATCTACCTCTATCTTC 645 Left TTAATTTTGGTTACATCCCTCTCTG 646 Right CTATCTTCTGTCCATTCTCTTCCAG 647 Left CAGACTCAGCTCAGTTAATTTTGGT 648 Right CTATCTTCTGTCCATTCTCTTCCAG 649 Left TATCCTGTTCCTCCCAGTTTAAGAT 650 Right CTATCTTCTGTCCATTCTCTTCCAG 651 Left TTAATTTTGGTTACATCCCTCTCTG 652 Right TCTATCTTCTGTCCATTCTCTTCCA 653 Left CAGACTCAGCTCAGTTAATTTTGGT 654 Right TCTATCTTCTGTCCATTCTCTTCCA 655 Left TTAATTTTGGTTACATCCCTCTCTG 656 Right CAGCCATCATCTACCTCTATCTTCT 657 Left TTAATTTTGGTTACATCCCTCTCTG 658 Right CTCAGCCATCATCTACCTCTATCTT 659 Left TTAATTTTGGTTACATCCCTCTCTG 660 Right AGCCATCATCTACCTCTATCTTCTG 661 Left CAGACTCAGCTCAGTTAATTTTGGT 662 Right CAGCCATCATCTACCTCTATCTTCT 663 Left CAGACTCAGCTCAGTTAATTTTGGT 664 Right CTCAGCCATCATCTACCTCTATCTT 665 Left CAGACTCAGCTCAGTTAATTTTGGT 666 Right AGCCATCATCTACCTCTATCTTCTG 667 Left TTAATTTTGGTTACATCCCTCTCTG 668 Right GCCATCATCTACCTCTATCTTCTGT 669 Left CAGACTCAGCTCAGTTAATTTTGGT 670 Right GCCATCATCTACCTCTATCTTCTGT 671 Left AGCTCAGTTAATTTTGGTTACATCC 672 Right TCAGCCATCATCTACCTCTATCTTC 673 Left AGCTCAGTTAATTTTGGTTACATCC 674 Right CTATCTTCTGTCCATTCTCTTCCAG 675 Left AGACTCAGCTCAGTTAATTTTGGTT 676 Right TCAGCCATCATCTACCTCTATCTTC 677 Left CTCAGCTCAGTTAATTTTGGTTACA 678 Right TCAGCCATCATCTACCTCTATCTTC 679 Left TATCCTGTTCCTCCCAGTTTAAGAT 680 Right TCTATCTTCTGTCCATTCTCTTCCA ALK Exon23 401-600 bases 681 Left TGTAGCTGCTGAAAATGTAACTTTG 682 Right TCAGCCATCATCTACCTCTATCTTC 683 Left TTAATTTTGGTTACATCCCTCTCTG 684 Right ACCTTCTGCAATGATTGTAAGTTTC 685 Left CAGACTCAGCTCAGTTAATTTTGGT 686 Right ACCTTCTGCAATGATTGTAAGTTTC 687 Left TTAATTTTGGTTACATCCCTCTCTG 688 Right CTTCTGCAATGATTGTAAGTTTCCT 689 Left CAGACTCAGCTCAGTTAATTTTGGT 690 Right CTTCTGCAATGATTGTAAGTTTCCT 691 Left TGTAGCTGCTGAAAATGTAACTTTG 692 Right CTATCTTCTGTCCATTCTCTTCCAG 693 Left TATCCTGTTCCTCCCAGTTTAAGAT 694 Right ACCTTCTGCAATGATTGTAAGTTTC 695 Left TATCCTGTTCCTCCCAGTTTAAGAT 696 Right CTTCTGCAATGATTGTAAGTTTCCT 697 Left TGTAGCTGCTGAAAATGTAACTTTG 698 Right TCTATCTTCTGTCCATTCTCTTCCA 699 Left TGTAGCTGCTGAAAATGTAACTTTG 700 Right CAGCCATCATCTACCTCTATCTTCT 701 Left TGTAGCTGCTGAAAATGTAACTTTG 702 Right AGCCATCATCTACCTCTATCTTCTG 703 Left TGTAGCTGCTGAAAATGTAACTTTG 704 Right CTCAGCCATCATCTACCTCTATCTT 705 Left TGTAGCTGCTGAAAATGTAACTTTG 706 Right GCCATCATCTACCTCTATCTTCTGT 707 Left TTAATTTTGGTTACATCCCTCTCTG 708 Right CACCTTCTGCAATGATTGTAAGTTT 709 Left AGCTCAGTTAATTTTGGTTACATCC 710 Right ACCTTCTGCAATGATTGTAAGTTTC 711 Left CAGACTCAGCTCAGTTAATTTTGGT 712 Right CACCTTCTGCAATGATTGTAAGTTT 713 Left AGCTCAGTTAATTTTGGTTACATCC 714 Right CTTCTGCAATGATTGTAAGTTTCCT 715 Left ATGCCTTTATACATTGTAGCTGCTG 716 Right TCAGCCATCATCTACCTCTATCTTC 717 Left TGTAGCTGCTGAAAATGTAACTTTG 718 Right GAGCCACTTAAATCTCTTTTCTTTG 719 Left TGTAGCTGCTGAAAATGTAACTTTG 720 Right TGAGCCACTTAAATCTCTTTTCTTT ALK Exon23 601-800 bases 721 Left TGTAGCTGCTGAAAATGTAACTTTG 722 Right ACCTTCTGCAATGATTGTAAGTTTC 723 Left TGTAGCTGCTGAAAATGTAACTTTG 724 Right GTTGAATTGTAATCCCTAGTGTTGG 725 Left TGTAGCTGCTGAAAATGTAACTTTG 726 Right CTTCTGCAATGATTGTAAGTTTCCT 727 Left TTAATTTTGGTTACATCCCTCTCTG 728 Right GCTATAGAATGTGGATATGGTTTGG 729 Left TTAATTTTGGTTACATCCCTCTCTG 730 Right GGCTATAGAATGTGGATATGGTTTG 731 Left CAGACTCAGCTCAGTTAATTTTGGT 732 Right GCTATAGAATGTGGATATGGTTTGG 733 Left CAGACTCAGCTCAGTTAATTTTGGT 734 Right GGCTATAGAATGTGGATATGGTTTG 735 Left TTAATTTTGGTTACATCCCTCTCTG 736 Right GTTGAATTGTAATCCCTAGTGTTGG 737 Left CAGACTCAGCTCAGTTAATTTTGGT 738 Right GTTGAATTGTAATCCCTAGTGTTGG 739 Left CCAGTATCTGAGAAAGACACTCTCC 740 Right TCAGCCATCATCTACCTCTATCTTC 741 Left TTAATTTTGGTTACATCCCTCTCTG 742 Right TAGAATGTGGATATGGTTTGGATTT 743 Left CAGACTCAGCTCAGTTAATTTTGGT 744 Right TAGAATGTGGATATGGTTTGGATTT 745 Left CCAGTATCTGAGAAAGACACTCTCC 746 Right CTATCTTCTGTCCATTCTCTTCCAG 747 Left TTAATTTTGGTTACATCCCTCTCTG 748 Right TCATGAGACCTGGTTGTTTAAAAGT 749 Left TTAATTTTGGTTACATCCCTCTCTG 750 Right CAGGCTATAGAATGTGGATATGGTT 751 Left TGTAGCTGCTGAAAATGTAACTTTG 752 Right CTCATGAGACCTGGTTGTTTAAAAG 753 Left CAGACTCAGCTCAGTTAATTTTGGT 754 Right TCATGAGACCTGGTTGTTTAAAAGT 755 Left CAGACTCAGCTCAGTTAATTTTGGT 756 Right CAGGCTATAGAATGTGGATATGGTT 757 Left TTAATTTTGGTTACATCCCTCTCTG 758 Right ATCCAGGCTATAGAATGTGGATATG 759 Left CAGACTCAGCTCAGTTAATTTTGGT 760 Right ATCCAGGCTATAGAATGTGGATATG ALK Exon23 801-1000 bases 761 Left TGTAGCTGCTGAAAATGTAACTTTG 762 Right GCTATAGAATGTGGATATGGTTTGG 763 Left TGTAGCTGCTGAAAATGTAACTTTG 764 Right GGCTATAGAATGTGGATATGGTTTG 765 Left TGTAGCTGCTGAAAATGTAACTTTG 766 Right GTCATGAAAGTTCTCCTCTGTGTTT 767 Left TGTAGCTGCTGAAAATGTAACTTTG 768 Right ATGAAAGTTCTCCTCTGTGTTTGTC 769 Left CCAGTATCTGAGAAAGACACTCTCC 770 Right ACCTTCTGCAATGATTGTAAGTTTC 771 Left TGTAGCTGCTGAAAATGTAACTTTG 772 Right TAGAATGTGGATATGGTTTGGATTT 773 Left TGTAGCTGCTGAAAATGTAACTTTG 774 Right CTCTAGTTTGGTTTTCCAGAGTCAG 775 Left TGTAGCTGCTGAAAATGTAACTTTG 776 Right CCTCTGTGTTTGTCTCTAGTTTGGT 777 Left CCAGTATCTGAGAAAGACACTCTCC 778 Right CTTCTGCAATGATTGTAAGTTTCCT 779 Left TGTAGCTGCTGAAAATGTAACTTTG 780 Right GAAAGTTCTCCTCTGTGTTTGTCTC 781 Left CATGTTAACAAGAAAACCCAAGTCT 782 Right TCAGCCATCATCTACCTCTATCTTC 783 Left TTAATTTTGGTTACATCCCTCTCTG 784 Right TATTATCCCTACTTGAGACGTGAGG 785 Left TTAATTTTGGTTACATCCCTCTCTG 786 Right CAGGTCAGTTGCTTGAGTAGTTACA 787 Left TTAATTTTGGTTACATCCCTCTCTG 788 Right GTCATGAAAGTTCTCCTCTGTGTTT 789 Left TTAATTTTGGTTACATCCCTCTCTG 790 Right ATGAAAGTTCTCCTCTGTGTTTGTC 791 Left CAGACTCAGCTCAGTTAATTTTGGT 792 Right TATTATCCCTACTTGAGACGTGAGG 793 Left CAGACTCAGCTCAGTTAATTTTGGT 794 Right CAGGTCAGTTGCTTGAGTAGTTACA 795 Left CAGACTCAGCTCAGTTAATTTTGGT 796 Right GTCATGAAAGTTCTCCTCTGTGTTT 797 Left CAGACTCAGCTCAGTTAATTTTGGT 798 Right ATGAAAGTTCTCCTCTGTGTTTGTC 799 Left TTAATTTTGGTTACATCCCTCTCTG 800 Right CTCTAGTTTGGTTTTCCAGAGTCAG ALK Exon23 2 kb 801 Left TTCTAACAGATCGATATCTCCAAGG 802 Right GCTATAGAATGTGGATATGGTTTGG 803 Left GAGCATGCTAGACTTTGACAGTACA 804 Right GCTATAGAATGTGGATATGGTTTGG 805 Left GAGCATGCTAGACTTTGACAGTACA 806 Right GGCTATAGAATGTGGATATGGTTTG 807 Left TTCTAACAGATCGATATCTCCAAGG 808 Right ACCTTCTGCAATGATTGTAAGTTTC 809 Left GAGCATGCTAGACTTTGACAGTACA 810 Right CCAAGCTCTGTTAACCATAAGATGT 811 Left TTCTAACAGATCGATATCTCCAAGG 812 Right GTTGAATTGTAATCCCTAGTGTTGG 813 Left TTCTAACAGATCGATATCTCCAAGG 814 Right CTTCTGCAATGATTGTAAGTTTCCT 815 Left GAGCATGCTAGACTTTGACAGTACA 816 Right GTTGAATTGTAATCCCTAGTGTTGG 817 Left ACAGGAGGACACACAAAATAACATT 818 Right ATAGTACAGTGGTTCGTTGAGGAAG 819 Left GAGCATGCTAGACTTTGACAGTACA 820 Right TATTATCCCTACTTGAGACGTGAGG 821 Left GAGCATGCTAGACTTTGACAGTACA 822 Right CAGGTCAGTTGCTTGAGTAGTTACA 823 Left GAGCATGCTAGACTTTGACAGTACA 824 Right GTCATGAAAGTTCTCCTCTGTGTTT 825 Left AGATATTTGACCTCAAGATCAGGTG 826 Right AGGTTAAAGGTTTAAGACTGCCCTA 827 Left CAGTAGGAGACTCCCAAACATTCTA 828 Right GTTGAATTGTAATCCCTAGTGTTGG 829 Left ACAGGAGGACACACAAAATAACATT 830 Right AGGTTAAAGGTTTAAGACTGCCCTA 831 Left AGATATTTGACCTCAAGATCAGGTG 832 Right CCAAGCTCTGTTAACCATAAGATGT 833 Left TTCTAACAGATCGATATCTCCAAGG 834 Right TAGAATGTGGATATGGTTTGGATTT 835 Left CATGTTAACAAGAAAACCCAAGTCT 836 Right ATAGTACAGTGGTTCGTTGAGGAAG 837 Left AGATATTTGACCTCAAGATCAGGTG 838 Right CACCTTTGAGATGTTCTAGTCCAAT 839 Left ACAGGAGGACACACAAAATAACATT 840 Right CACCTTTGAGATGTTCTAGTCCAAT ALK Exon21-23 2 kb 841 Left TTGACTCTGTCTCCTCTTGTCTTCT 842 Right TCAGCCATCATCTACCTCTATCTTC 843 Left TTGACTCTGTCTCCTCTTGTCTTCT 844 Right CTATCTTCTGTCCATTCTCTTCCAG 845 Left TTTGACTCTGTCTCCTCTTGTCTTC 846 Right TCAGCCATCATCTACCTCTATCTTC 847 Left ACTCTGTCTCCTCTTGTCTTCTCCT 848 Right TCAGCCATCATCTACCTCTATCTTC 849 Left GTTTGACTCTGTCTCCTCTTGTCTT 850 Right TCAGCCATCATCTACCTCTATCTTC 851 Left TTTGACTCTGTCTCCTCTTGTCTTC 852 Right CTATCTTCTGTCCATTCTCTTCCAG 853 Left ACTCTGTCTCCTCTTGTCTTCTCCT 854 Right CTATCTTCTGTCCATTCTCTTCCAG 855 Left GTTTGACTCTGTCTCCTCTTGTCTT 856 Right CTATCTTCTGTCCATTCTCTTCCAG 857 Left TTGACTCTGTCTCCTCTTGTCTTCT 858 Right TCTATCTTCTGTCCATTCTCTTCCA 859 Left TGACTCTGTCTCCTCTTGTCTTCTC 860 Right TCAGCCATCATCTACCTCTATCTTC 861 Left TTGACTCTGTCTCCTCTTGTCTTCT 862 Right CAGCCATCATCTACCTCTATCTTCT 863 Left TTGACTCTGTCTCCTCTTGTCTTCT 864 Right CTCAGCCATCATCTACCTCTATCTT 865 Left TTGACTCTGTCTCCTCTTGTCTTCT 866 Right AGCCATCATCTACCTCTATCTTCTG 867 Left TGACTCTGTCTCCTCTTGTCTTCTC 868 Right CTATCTTCTGTCCATTCTCTTCCAG 869 Left TTGACTCTGTCTCCTCTTGTCTTCT 870 Right GCCATCATCTACCTCTATCTTCTGT 871 Left TGTTTGACTCTGTCTCCTCTTGTCT 872 Right TCAGCCATCATCTACCTCTATCTTC 873 Left CTGTTTGACTCTGTCTCCTCTTGTC 874 Right TCAGCCATCATCTACCTCTATCTTC 875 Left CTCTGTCTCCTCTTGTCTTCTCCTT 876 Right TCAGCCATCATCTACCTCTATCTTC 877 Left TTTGACTCTGTCTCCTCTTGTCTTC 878 Right TCTATCTTCTGTCCATTCTCTTCCA 879 Left TGTTTGACTCTGTCTCCTCTTGTCT 880 Right CTATCTTCTGTCCATTCTCTTCCAG ALK Exon21-23 5 kb 881 Left CAGTGTAGGGGCTGAATGTTATC 882 Right TACAACTTTCTCTCCTTAAGCCTCA 883 Left CAGTGTAGGGGCTGAATGTTATC 884 Right ACAACTTTCTCTCCTTAAGCCTCA 885 Left CAGTGTAGGGGCTGAATGTTATC 886 Right ATACAACTTTCTCTCCTTAAGCCTCA 887 Left GCAGTGTAGGGGCTGAATGTTAT 888 Right TACAACTTTCTCTCCTTAAGCCTCA 889 Left CAGACTCCTCTAGCCACAAAAGG 890 Right TACAACTTTCTCTCCTTAAGCCTCA 891 Left GCAGACTCCTCTAGCCACAAAAG 892 Right TACAACTTTCTCTCCTTAAGCCTCA 893 Left GCAGACTCCTCTAGCCACAAAA 894 Right TACAACTTTCTCTCCTTAAGCCTCA 895 Left AGACTCCTCTAGCCACAAAAGG 896 Right TACAACTTTCTCTCCTTAAGCCTCA 897 Left CAGTGTAGGGGCTGAATGTTATC 898 Right GCATGCATACAACTTTCTCTCCTT 899 Left GCAGTGTAGGGGCTGAATGTTATC 900 Right TACAACTTTCTCTCCTTAAGCCTCA 901 Left GTAGGGGCTGAATGTTATCACAGC 902 Right TACAACTTTCTCTCCTTAAGCCTCA 903 Left GAGGACAAGCCTTGACATTCAG 904 Right TACAACTTTCTCTCCTTAAGCCTCA 905 Left ATGTTGGCTTACATTAACTCCCATA 906 Right TTTGCAAAGTCCCTCTCCTTT 907 Left GTTATCACAGCACCGCAGACT 908 Right TACAACTTTCTCTCCTTAAGCCTCA 909 Left GCAGACTCCTCTAGCCACAAA 910 Right TACAACTTTCTCTCCTTAAGCCTCA 911 Left GCAGTGTAGGGGCTGAATGTTA 912 Right TACAACTTTCTCTCCTTAAGCCTCA 913 Left TAGGGGCTGAATGTTATCACAGC 914 Right TACAACTTTCTCTCCTTAAGCCTCA 915 Left CAGACTCCTCTAGCCACAAAAGG 916 Right GCATGCATACAACTTTCTCTCCTTA 917 Left GCAGTGTAGGGGCTGAATGTTAT 918 Right ACAACTTTCTCTCCTTAAGCCTCA 919 Left ATGTTGGCTTACATTAACTCCCATA 920 Right CAAAGTCCCTCTCCTTTGCAT ALK Exon24 130-150 bases 921 Left AGTGGCCCGCTTCTGTCT 922 Right GATGACAGGAAGAGCACAGTCAC 923 Left CGCTTCTGTCTCCCCACAG 924 Right GATGACAGGAAGAGCACAGTCAC 925 Left CGCTTCTGTCTCCCCACA 926 Right GATGACAGGAAGAGCACAGTCAC 927 Left AGTGGCCCGCTTCTGTCT 928 Right AGGATGACAGGAAGAGCACAGT 929 Left CGCTTCTGTCTCCCCACAG 930 Right AGGATGACAGGAAGAGCACAGT 931 Left CGCTTCTGTCTCCCCACA 932 Right AGGATGACAGGAAGAGCACAGT 933 Left AGTGGCCCGCTTCTGTCT 934 Right GATGACAGGAAGAGCACAGTCA 935 Left CGCTTCTGTCTCCCCACAG 936 Right GATGACAGGAAGAGCACAGTCA 937 Left CGCTTCTGTCTCCCCACA 938 Right GATGACAGGAAGAGCACAGTCA 939 Left AGTGGCCCGCTTCTGTCT 940 Right ATGACAGGAAGAGCACAGTCAC 941 Left CGCTTCTGTCTCCCCACAG 942 Right ATGACAGGAAGAGCACAGTCAC 943 Left CGCTTCTGTCTCCCCACA 944 Right ATGACAGGAAGAGCACAGTCAC 945 Left AGTGGCCCGCTTCTGTCT 946 Right AGGATGACAGGAAGAGCACAGTC 947 Left CGCTTCTGTCTCCCCACAG 948 Right AGGATGACAGGAAGAGCACAGTC 949 Left CGCTTCTGTCTCCCCACA 950 Right AGGATGACAGGAAGAGCACAGTC 951 Left AGTGGCCCGCTTCTGTCT 952 Right GGATGACAGGAAGAGCACAGTC 953 Left CGCTTCTGTCTCCCCACAG 954 Right GGATGACAGGAAGAGCACAGTC 955 Left CGCTTCTGTCTCCCCACAG 956 Right GACAGGATGACAGGAAGAGCAC 957 Left CGCTTCTGTCTCCCCACA 958 Right GGATGACAGGAAGAGCACAGTC 959 Left CGCTTCTGTCTCCCCACA 960 Right GACAGGATGACAGGAAGAGCAC ALK Exon24 161-200 bases 961 Left ATTTCAGATTTCCCTCCTCTCACT 962 Right GATGACAGGAAGAGCACAGTCAC 963 Left ATTTCAGATTTCCCTCCTCTCACT 964 Right AGGATGACAGGAAGAGCACAGT 965 Left ATTTCAGATTTCCCTCCTCTCACT 966 Right GATGACAGGAAGAGCACAGTCA 967 Left ATTTCAGATTTCCCTCCTCTCAC 968 Right GATGACAGGAAGAGCACAGTCAC 969 Left ATTTCAGATTTCCCTCCTCTCAC 970 Right AGGATGACAGGAAGAGCACAGT 971 Left ATTTCAGATTTCCCTCCTCTCACT 972 Right ATGACAGGAAGAGCACAGTCAC 973 Left ATTTCAGATTTCCCTCCTCTCAC 974 Right GATGACAGGAAGAGCACAGTCA 975 Left ATTTCAGATTTCCCTCCTCTCAC 976 Right ATGACAGGAAGAGCACAGTCAC 977 Left TTTCAGATTTCCCTCCTCTCACT 978 Right GATGACAGGAAGAGCACAGTCAC 979 Left TTTCAGATTTCCCTCCTCTCACT 980 Right AGGATGACAGGAAGAGCACAGT 981 Left ATTTCAGATTTCCCTCCTCTCACT 982 Right AGGATGACAGGAAGAGCACAGTC 983 Left ATTTCAGATTTCCCTCCTCTCACT 984 Right GGATGACAGGAAGAGCACAGTC 985 Left TTTCAGATTTCCCTCCTCTCACT 986 Right GATGACAGGAAGAGCACAGTCA 987 Left ATTTCAGATTTCCCTCCTCTCAC 988 Right AGGATGACAGGAAGAGCACAGTC 989 Left TTTCAGATTTCCCTCCTCTCACT 990 Right ATGACAGGAAGAGCACAGTCAC 991 Left ATTTCAGATTTCCCTCCTCTCAC 992 Right GGATGACAGGAAGAGCACAGTC 993 Left ATTTCAGATTTCCCTCCTCTCACT 994 Right AGGATGACAGGAAGAGCACAG 995 Left ATTTCCCTCCTCTCACTGACAA 996 Right GATGACAGGAAGAGCACAGTCAC 997 Left ATTTCCCTCCTCTCACTGACAA 998 Right AGGATGACAGGAAGAGCACAGT 999 Left CATTTCAGATTTCCCTCCTCTCACT 1000 Right GATGACAGGAAGAGCACAGTCAC ALK Exon24 201-300 bases 1001 Left ATTTCAGATTTCCCTCCTCTCACT 1002 Right GAGACCTAGTATTCTGCTCTGAAGG 1003 Left ATTTCAGATTTCCCTCCTCTCACT 1004 Right GGAGACCTAGTATTCTGCTCTGAAG 1005 Left ATTTCAGATTTCCCTCCTCTCACT 1006 Right CTCTGGAGGGAGACCTAGTATTCTG 1007 Left ATTTCAGATTTCCCTCCTCTCAC 1008 Right GAGACCTAGTATTCTGCTCTGAAGG 1009 Left ATTTCAGATTTCCCTCCTCTCAC 1010 Right GGAGACCTAGTATTCTGCTCTGAAG 1011 Left ATTTCAGATTTCCCTCCTCTCACT 1012 Right AGGGAGACCTAGTATTCTGCTCTGA 1013 Left ATTTCAGATTTCCCTCCTCTCACT 1014 Right TCTGGAGGGAGACCTAGTATTCTG 1015 Left ATTTCAGATTTCCCTCCTCTCAC 1016 Right CTCTGGAGGGAGACCTAGTATTCTG 1017 Left ATTTCAGATTTCCCTCCTCTCACT 1018 Right GGGAGACCTAGTATTCTGCTCTGA 1019 Left ATTTCAGATTTCCCTCCTCTCAC 1020 Right AGGGAGACCTAGTATTCTGCTCTGA 1021 Left ATTTCAGATTTCCCTCCTCTCAC 1022 Right TCTGGAGGGAGACCTAGTATTCTG 1023 Left CTCCTCTCACTGACAAGCTCCT 1024 Right AAACAAAGCTGAATCATCCTACATC 1025 Left ATTTCAGATTTCCCTCCTCTCAC 1026 Right GGGAGACCTAGTATTCTGCTCTGA 1027 Left TTTCAGATTTCCCTCCTCTCACT 1028 Right GAGACCTAGTATTCTGCTCTGAAGG 1029 Left TTTCAGATTTCCCTCCTCTCACT 1030 Right GGAGACCTAGTATTCTGCTCTGAAG 1031 Left TTTCAGATTTCCCTCCTCTCACT 1032 Right CTCTGGAGGGAGACCTAGTATTCTG 1033 Left TTTCAGATTTCCCTCCTCTCACT 1034 Right AGGGAGACCTAGTATTCTGCTCTGA 1035 Left TTTCAGATTTCCCTCCTCTCACT 1036 Right TCTGGAGGGAGACCTAGTATTCTG 1037 Left ATTTCAGATTTCCCTCCTCTCACT 1038 Right GCTCTGGAGGGAGACCTAGTATTC 1039 Left TTTCAGATTTCCCTCCTCTCACT 1040 Right GGGAGACCTAGTATTCTGCTCTGA ALK Exon24 301-400 bases 1041 Left ATTTCAGATTTCCCTCCTCTCACT 1042 Right AAACAAAGCTGAATCATCCTACATC 1043 Left ATTTCAGATTTCCCTCCTCTCACT 1044 Right ACAATAAAACAAAGCTGAATCATCC 1045 Left ATTTCAGATTTCCCTCCTCTCAC 1046 Right AAACAAAGCTGAATCATCCTACATC 1047 Left ATTTCAGATTTCCCTCCTCTCAC 1048 Right ACAATAAAACAAAGCTGAATCATCC 1049 Left ATTTCAGATTTCCCTCCTCTCACT 1050 Right AAAACAAAGCTGAATCATCCTACAT 1051 Left ATTTCAGATTTCCCTCCTCTCAC 1052 Right AAAACAAAGCTGAATCATCCTACAT 1053 Left ATTTCAGATTTCCCTCCTCTCACT 1054 Right TAAAACAAAGCTGAATCATCCTACA 1055 Left TTTCAGATTTCCCTCCTCTCACT 1056 Right AAACAAAGCTGAATCATCCTACATC 1057 Left ATTTCAGATTTCCCTCCTCTCAC 1058 Right TAAAACAAAGCTGAATCATCCTACA 1059 Left TTTCAGATTTCCCTCCTCTCACT 1060 Right ACAATAAAACAAAGCTGAATCATCC 1061 Left ATTTCAGATTTCCCTCCTCTCACT 1062 Right CAATAAAACAAAGCTGAATCATCCTA 1063 Left ATTTCAGATTTCCCTCCTCTCACT 1064 Right AGCTGAATCATCCTACATCCAAAT 1065 Left AATGCCTCCAGGTGATTTCTAAT 1066 Right GAGACCTAGTATTCTGCTCTGAAGG 1067 Left AATGCCTCCAGGTGATTTCTAAT 1068 Right GGAGACCTAGTATTCTGCTCTGAAG 1069 Left TTTCAGATTTCCCTCCTCTCACT 1070 Right AAAACAAAGCTGAATCATCCTACAT 1071 Left ATTTCAGATTTCCCTCCTCTCAC 1072 Right CAATAAAACAAAGCTGAATCATCCTA 1073 Left ATTTCAGATTTCCCTCCTCTCACT 1074 Right AAGCTGAATCATCCTACATCCAAAT 1075 Left AAATGCCTCCAGGTGATTTCTAAT 1076 Right GAGACCTAGTATTCTGCTCTGAAGG 1077 Left AAATGCCTCCAGGTGATTTCTAAT 1078 Right GGAGACCTAGTATTCTGCTCTGAAG 1079 Left AATGCCTCCAGGTGATTTCTAAT 1080 Right CTCTGGAGGGAGACCTAGTATTCTG ALK Exon24 401-600 bases 1081 Left TGCACAATAAATTAAAAGGGAAAGA 1082 Right AAACAAAGCTGAATCATCCTACATC 1083 Left TGCACAATAAATTAAAAGGGAAAGA 1084 Right ACAATAAAACAAAGCTGAATCATCC 1085 Left GTGCACAATAAATTAAAAGGGAAAG 1086 Right AAACAAAGCTGAATCATCCTACATC 1087 Left TGCACAATAAATTAAAAGGGAAAGA 1088 Right AAAACAAAGCTGAATCATCCTACAT 1089 Left ATCATATTACCTGGGAAGACTTCAA 1090 Right AAACAAAGCTGAATCATCCTACATC 1091 Left GTGCACAATAAATTAAAAGGGAAAG 1092 Right ACAATAAAACAAAGCTGAATCATCC 1093 Left TGTGCACAATAAATTAAAAGGGAAA 1094 Right AAACAAAGCTGAATCATCCTACATC 1095 Left TGCACAATAAATTAAAAGGGAAAGA 1096 Right TAAAACAAAGCTGAATCATCCTACA 1097 Left TAAATTAAAAGGGAAAGAACACCTG 1098 Right AAACAAAGCTGAATCATCCTACATC 1099 Left GCACAATAAATTAAAAGGGAAAGAA 1100 Right AAACAAAGCTGAATCATCCTACATC 1101 Left TGGGAAGACTTCAAATGTACAAATA 1102 Right AAACAAAGCTGAATCATCCTACATC 1103 Left TGCACAATAAATTAAAAGGGAAAGA 1104 Right GAGACCTAGTATTCTGCTCTGAAGG 1105 Left TGCACAATAAATTAAAAGGGAAAGA 1106 Right GGAGACCTAGTATTCTGCTCTGAAG 1107 Left TGTGCACAATAAATTAAAAGGGAAA 1108 Right ACAATAAAACAAAGCTGAATCATCC 1109 Left TGTTTATAAATTGGGGGTATTCAAA 1110 Right GAGACCTAGTATTCTGCTCTGAAGG 1111 Left TTGTTTATAAATTGGGGGTATTCAA 1112 Right GAGACCTAGTATTCTGCTCTGAAGG 1113 Left TGTTTATAAATTGGGGGTATTCAAA 1114 Right GGAGACCTAGTATTCTGCTCTGAAG 1115 Left TTGTTTATAAATTGGGGGTATTCAA 1116 Right GGAGACCTAGTATTCTGCTCTGAAG 1117 Left GTGCACAATAAATTAAAAGGGAAAG 1118 Right AAAACAAAGCTGAATCATCCTACAT 1119 Left TAAATTAAAAGGGAAAGAACACCTG 1120 Right ACAATAAAACAAAGCTGAATCATCC ALK Exon24 601-800 bases 1121 Left TGTTTATAAATTGGGGGTATTCAAA 1122 Right AAACAAAGCTGAATCATCCTACATC 1123 Left TTGTTTATAAATTGGGGGTATTCAA 1124 Right AAACAAAGCTGAATCATCCTACATC 1125 Left TGTTTATAAATTGGGGGTATTCAAA 1126 Right ACAATAAAACAAAGCTGAATCATCC 1127 Left TGCACAATAAATTAAAAGGGAAAGA 1128 Right AGTTACCATCTCAAAGACAAAGCTG 1129 Left TGTTTATAAATTGGGGGTATTCAAA 1130 Right AGTTACCATCTCAAAGACAAAGCTG 1131 Left TTGTTTATAAATTGGGGGTATTCAA 1132 Right AGTTACCATCTCAAAGACAAAGCTG 1133 Left CAAACTTGTTTATAAATTGGGGGTA 1134 Right AAACAAAGCTGAATCATCCTACATC 1135 Left CTTGTTTATAAATTGGGGGTATTCA 1136 Right AAACAAAGCTGAATCATCCTACATC 1137 Left TGCACAATAAATTAAAAGGGAAAGA 1138 Right GCAAGTGAATCCCTGATAGAATAAG 1139 Left CTGGATCTGCTTGAAGAAAATTAGT 1140 Right AAACAAAGCTGAATCATCCTACATC 1141 Left CAAACTTGTTTATAAATTGGGGGTA 1142 Right ACAATAAAACAAAGCTGAATCATCC 1143 Left TTTATAAATTGGGGGTATTCAAATG 1144 Right AAACAAAGCTGAATCATCCTACATC 1145 Left TGTTTATAAATTGGGGGTATTCAAA 1146 Right AAAACAAAGCTGAATCATCCTACAT 1147 Left TTGTTTATAAATTGGGGGTATTCAA 1148 Right AAAACAAAGCTGAATCATCCTACAT 1149 Left CTGGATCTGCTTGAAGAAAATTAGT 1150 Right ACAATAAAACAAAGCTGAATCATCC 1151 Left GTGCACAATAAATTAAAAGGGAAAG 1152 Right AGTTACCATCTCAAAGACAAAGCTG 1153 Left ATCATATTACCTGGGAAGACTTCAA 1154 Right GCGAGGATATTTTATGACACTTGTT 1155 Left TTTATAAATTGGGGGTATTCAAATG 1156 Right ACAATAAAACAAAGCTGAATCATCC 1157 Left ATCTCCTTTTGAATGAAAGAGACCT 1158 Right GAGACCTAGTATTCTGCTCTGAAGG 1159 Left ATCTCCTTTTGAATGAAAGAGACCT 1160 Right GGAGACCTAGTATTCTGCTCTGAAG ALK Exon24 801-1000 bases 1161 Left TAGGAATTAAAAGAGAGGCCAAGAT 1162 Right AAACAAAGCTGAATCATCCTACATC 1163 Left ATCTCCTTTTGAATGAAAGAGACCT 1164 Right AAACAAAGCTGAATCATCCTACATC 1165 Left TAGGAATTAAAAGAGAGGCCAAGAT 1166 Right ACAATAAAACAAAGCTGAATCATCC 1167 Left ATCTCCTTTTGAATGAAAGAGACCT 1168 Right ACAATAAAACAAAGCTGAATCATCC 1169 Left TGCACAATAAATTAAAAGGGAAAGA 1170 Right AAAGATGACTAAAACAGCATCCTTG 1171 Left ACGTCAGGGATTTAGGAATTAAAAG 1172 Right ACAATAAAACAAAGCTGAATCATCC 1173 Left TGCACAATAAATTAAAAGGGAAAGA 1174 Right GCGAGGATATTTTATGACACTTGTT 1175 Left TGTTTATAAATTGGGGGTATTCAAA 1176 Right GCGAGGATATTTTATGACACTTGTT 1177 Left TTGTTTATAAATTGGGGGTATTCAA 1178 Right GCGAGGATATTTTATGACACTTGTT 1179 Left TGCACAATAAATTAAAAGGGAAAGA 1180 Right TTGTCAAAAATGCAATTCCTTAACT 1181 Left TTTAGGAATTAAAAGAGAGGCCAAG 1182 Right AAACAAAGCTGAATCATCCTACATC 1183 Left TAGGAATTAAAAGAGAGGCCAAGAT 1184 Right AAAACAAAGCTGAATCATCCTACAT 1185 Left ATCTCCTTTTGAATGAAAGAGACCT 1186 Right AAAACAAAGCTGAATCATCCTACAT 1187 Left AAAGCTTGAGATAGCTCATAATTGC 1188 Right AAACAAAGCTGAATCATCCTACATC 1189 Left TTTAGGAATTAAAAGAGAGGCCAAG 1190 Right ACAATAAAACAAAGCTGAATCATCC 1191 Left ACGTCAGGGATTTAGGAATTAAAAG 1192 Right AAAACAAAGCTGAATCATCCTACAT 1193 Left TGTTTATAAATTGGGGGTATTCAAA 1194 Right GCAAGTGAATCCCTGATAGAATAAG 1195 Left TTGTTTATAAATTGGGGGTATTCAA 1196 Right GCAAGTGAATCCCTGATAGAATAAG 1197 Left GTGCACAATAAATTAAAAGGGAAAG 1198 Right AAAGATGACTAAAACAGCATCCTTG 1199 Left CAAACTTGTTTATAAATTGGGGGTA 1200 Right GCGAGGATATTTTATGACACTTGTT ALK Exon24 2 kb 1201 Left ATCTCCTTTTGAATGAAAGAGACCT 1202 Right CATGTTAGGAGTGACTTTGGAACTT 1203 Left ACTGTAGTCACATACATACGCTCCA 1204 Right AAACAAAGCTGAATCATCCTACATC 1205 Left ATCTCCTTTTGAATGAAAGAGACCT 1206 Right CTAAAAGGATGAAGTGACAGGAAGA 1207 Left ATCTCCTTTTGAATGAAAGAGACCT 1208 Right TAAAAGGATGAAGTGACAGGAAGAG 1209 Left ACTGTAGTCACATACATACGCTCCA 1210 Right GCGAGGATATTTTATGACACTTGTT 1211 Left ACTGTAGTCACATACATACGCTCCA 1212 Right ACAATAAAACAAAGCTGAATCATCC 1213 Left ACGTCAGGGATTTAGGAATTAAAAG 1214 Right CAAGTGTACTTCCTGACCTCTCATT 1215 Left TGCACAATAAATTAAAAGGGAAAGA 1216 Right CATGTTAGGAGTGACTTTGGAACTT 1217 Left ACTGTAGTCACATACATACGCTCCA 1218 Right AGTTACCATCTCAAAGACAAAGCTG 1219 Left TGTTTATAAATTGGGGGTATTCAAA 1220 Right CATGTTAGGAGTGACTTTGGAACTT 1221 Left TTGTTTATAAATTGGGGGTATTCAA 1222 Right CATGTTAGGAGTGACTTTGGAACTT 1223 Left ATCTCCTTTTGAATGAAAGAGACCT 1224 Right ATAAGCTCTTCTGAGAGTTGACTGC 1225 Left AGGCACTCTCTCTTCCATTTTAACT 1226 Right AAACAAAGCTGAATCATCCTACATC 1227 Left TGCACAATAAATTAAAAGGGAAAGA 1228 Right AAGGGCTGAAAAACTACCTTAAAAA 1229 Left TGCACAATAAATTAAAAGGGAAAGA 1230 Right AAAGGGCTGAAAAACTACCTTAAAA 1231 Left TGCACAATAAATTAAAAGGGAAAGA 1232 Right TTAGAAGGAGCAGATGGTAAAGCTA 1233 Left TGTTTATAAATTGGGGGTATTCAAA 1234 Right AAGGGCTGAAAAACTACCTTAAAAA 1235 Left TGTTTATAAATTGGGGGTATTCAAA 1236 Right AAAGGGCTGAAAAACTACCTTAAAA 1237 Left TTGTTTATAAATTGGGGGTATTCAA 1238 Right AAAGGGCTGAAAAACTACCTTAAAA 1239 Left TTGTTTATAAATTGGGGGTATTCAA 1240 Right AAGGGCTGAAAAACTACCTTAAAAA ALK Exon25 130-160 bases 1241 Left TCCTAGGGATAAAATTAGGAAATGC 1242 Right AGGGGTGAGGCAGTCTTTACTC 1243 Left TCCTAGGGATAAAATTAGGAAATGC 1244 Right GAGGGGTGAGGCAGTCTTTACT 1245 Left TCCTAGGGATAAAATTAGGAAATGC 1246 Right GGGGTGAGGCAGTCTTTACTC 1247 Left TCCTAGGGATAAAATTAGGAAATGC 1248 Right GAGGGGTGAGGCAGTCTTTAC 1249 Left TCCTAGGGATAAAATTAGGAAATGC 1250 Right GGGTGAGGCAGTCTTTACTCA 1251 Left TCCTAGGGATAAAATTAGGAAATGC 1252 Right GAGGGGTGAGGCAGTCTTTACTC 1253 Left TCCTAGGGATAAAATTAGGAAATGC 1254 Right AGGGGTGAGGCAGTCTTTACT 1255 Left TCCTAGGGATAAAATTAGGAAATGC 1256 Right AGGGGTGAGGCAGTCTTTACTCA 1257 Left TCCTAGGGATAAAATTAGGAAATGC 1258 Right GAGGGGTGAGGCAGTCTTTA 1259 Left TTCCTAGGGATAAAATTAGGAAATG 1260 Right AGGGGTGAGGCAGTCTTTACTC 1261 Left TCCTAGGGATAAAATTAGGAAATGC 1262 Right GGGGTGAGGCAGTCTTTACTCA 1263 Left TTCCTAGGGATAAAATTAGGAAATG 1264 Right GGGGTGAGGCAGTCTTTACTC 1265 Left TTCCTAGGGATAAAATTAGGAAATGC 1266 Right AGGGGTGAGGCAGTCTTTACTC 1267 Left CTTCCTAGGGATAAAATTAGGAAATG 1268 Right GGGGTGAGGCAGTCTTTACTC 1269 Left TCCTAGGGATAAAATTAGGAAATGC ALK Exon25 161-200 bases 1270 Right GAGGGGTGAGGCAGTCTTTACTCA 1271 Left TTCCTAGGGATAAAATTAGGAAATGC 1272 Right GGGGTGAGGCAGTCTTTACTC 1273 Left CCTAGGGATAAAATTAGGAAATGC 1274 Right AGGGGTGAGGCAGTCTTTACTC 1275 Left CCTAGGGATAAAATTAGGAAATGC 1276 Right GAGGGGTGAGGCAGTCTTTACT 1277 Left TTCCTAGGGATAAAATTAGGAAATG 1278 Right GGGTGAGGCAGTCTTTACTCA 1279 Left TTCCTAGGGATAAAATTAGGAAATG 1280 Right AGGGGTGAGGCAGTCTTTACT 1281 Left CTTGGAGATAAAATCCTAGTGATGG 1282 Right GGGGTGAGGCAGTCTTTACTC 1283 Left TCCTAGGGATAAAATTAGGAAATGC 1284 Right GCTGAGGTGGAAGAGACAGG 1285 Left TCCTAGGGATAAAATTAGGAAATGC 1286 Right GGCTGAGGTGGAAGAGACAG 1287 Left CTTGGAGATAAAATCCTAGTGATGG 1288 Right GGGTGAGGCAGTCTTTACTCA 1289 Left TGTACACTCATCTTCCTAGGGATAAA 1290 Right GAGGGGTGAGGCAGTCTTTACT 1291 Left TGTACACTCATCTTCCTAGGGATAAA 1292 Right GAGGGGTGAGGCAGTCTTTAC 1293 Left TCATCTTCCTAGGGATAAAATTAGG 1294 Right AGGGGTGAGGCAGTCTTTACTC 1295 Left TCATCTTCCTAGGGATAAAATTAGG 1296 Right GAGGGGTGAGGCAGTCTTTACT 1297 Left TTCCTAGGGATAAAATTAGGAAATG 1298 Right GAGGGGTGAGGCAGTCTTTACT 1299 Left TCATCTTCCTAGGGATAAAATTAGG 1300 Right GGGGTGAGGCAGTCTTTACTC 1301 Left TCATCTTCCTAGGGATAAAATTAGG 1302 Right GAGGGGTGAGGCAGTCTTTAC 1303 Left TTCCTAGGGATAAAATTAGGAAATG 1304 Right GAGGGGTGAGGCAGTCTTTAC 1305 Left CTCATCTTCCTAGGGATAAAATTAGG 1306 Right AGGGGTGAGGCAGTCTTTACTC 1307 Left CTCATCTTCCTAGGGATAAAATTAGG 1308 Right GAGGGGTGAGGCAGTCTTTACT 1309 Left CTTCCTAGGGATAAAATTAGGAAATG 1310 Right AGGGGTGAGGCAGTCTTTACTC 1311 Left CTTCCTAGGGATAAAATTAGGAAATG 1312 Right GAGGGGTGAGGCAGTCTTTACT 1313 Left CTCATCTTCCTAGGGATAAAATTAGG 1314 Right GGGGTGAGGCAGTCTTTACTC 1315 Left CTCATCTTCCTAGGGATAAAATTAGG 1316 Right GAGGGGTGAGGCAGTCTTTAC 1317 Left TTCCTAGGGATAAAATTAGGAAATGC 1318 Right GAGGGGTGAGGCAGTCTTTACT 1319 Left CTTCCTAGGGATAAAATTAGGAAATG 1320 Right GAGGGGTGAGGCAGTCTTTAC ALK Exon25 201-300 bases 1321 Left TCCTAGGGATAAAATTAGGAAATGC 1322 Right AGCCTGAAAAGGAACTTAGTGAAAT 1323 Left TCCTAGGGATAAAATTAGGAAATGC 1324 Right TAGCCTGAAAAGGAACTTAGTGAAA 1325 Left TCCTAGGGATAAAATTAGGAAATGC 1326 Right CATAGCCTGAAAAGGAACTTAGTGA 1327 Left TCCTAGGGATAAAATTAGGAAATGC 1328 Right CTAATTAAGGTTTCCCATAGCCTGA 1329 Left TCCTAGGGATAAAATTAGGAAATGC 1330 Right ATAGCCTGAAAAGGAACTTAGTGAAA 1331 Left TCCTAGGGATAAAATTAGGAAATGC 1332 Right TAGCCTGAAAAGGAACTTAGTGAAAT 1333 Left TCCTAGGGATAAAATTAGGAAATGC 1334 Right AGGTAGAAAGTTGACAGGGTACCAG 1335 Left TCCTAGGGATAAAATTAGGAAATGC 1336 Right AGCCTGAAAAGGAACTTAGTGAAA 1337 Left TCCTAGGGATAAAATTAGGAAATGC 1338 Right TAATTAAGGTTTCCCATAGCCTGA 1339 Left TCCTAGGGATAAAATTAGGAAATGC 1340 Right TAATTAAGGTTTCCCATAGCCTGAA 1341 Left TCCTAGGGATAAAATTAGGAAATGC 1342 Right GGTAGAAAGTTGACAGGGTACCAG 1343 Left TGTACACTCATCTTCCTAGGGATAAA 1344 Right AGCCTGAAAAGGAACTTAGTGAAAT 1345 Left TGTACACTCATCTTCCTAGGGATAAA 1346 Right TAGCCTGAAAAGGAACTTAGTGAAA 1347 Left TCCTAGGGATAAAATTAGGAAATGC 1348 Right CCATAGCCTGAAAAGGAACTTAGTG 1349 Left CTTGGAGATAAAATCCTAGTGATGG 1350 Right AGGTAGAAAGTTGACAGGGTACCAG 1351 Left TGTACACTCATCTTCCTAGGGATAAA 1352 Right CATAGCCTGAAAAGGAACTTAGTGA 1353 Left TCCTAGGGATAAAATTAGGAAATGC 1354 Right CACTAATTAAGGTTTCCCATAGCC 1355 Left TCATCTTCCTAGGGATAAAATTAGG 1356 Right AGCCTGAAAAGGAACTTAGTGAAAT 1357 Left TCCTAGGGATAAAATTAGGAAATGC 1358 Right GCCTGAAAAGGAACTTAGTGAAAT 1359 Left AACTTCAGCTTGGAGATAAAATCCT 1360 Right ACAGGGTACCAGGAGATGATGTAAG ALK Exon25 301-400 bases 1361 Left AACTTCAGCTTGGAGATAAAATCCT 1362 Right TGGTCACTAATTAAGGTTTCCCATA 1363 Left GGGAAGGAACTATTTAAACTTCAGC 1364 Right TGGTCACTAATTAAGGTTTCCCATA 1365 Left TCCTAGGGATAAAATTAGGAAATGC 1366 Right TGGTCACTAATTAAGGTTTCCCATA 1367 Left AACTTCAGCTTGGAGATAAAATCCT 1368 Right AGCCTGAAAAGGAACTTAGTGAAAT 1369 Left GGGAAGGAACTATTTAAACTTCAGC 1370 Right AGCCTGAAAAGGAACTTAGTGAAAT 1371 Left AACTTCAGCTTGGAGATAAAATCCT 1372 Right TAGCCTGAAAAGGAACTTAGTGAAA 1373 Left GGGAAGGAACTATTTAAACTTCAGC 1374 Right TAGCCTGAAAAGGAACTTAGTGAAA 1375 Left AACTTCAGCTTGGAGATAAAATCCT 1376 Right CATAGCCTGAAAAGGAACTTAGTGA 1377 Left GGGAAGGAACTATTTAAACTTCAGC 1378 Right CATAGCCTGAAAAGGAACTTAGTGA 1379 Left AAACTTCAGCTTGGAGATAAAATCC 1380 Right TGGTCACTAATTAAGGTTTCCCATA 1381 Left AAACTTCAGCTTGGAGATAAAATCC 1382 Right AGCCTGAAAAGGAACTTAGTGAAAT 1383 Left AAACTTCAGCTTGGAGATAAAATCC 1384 Right TAGCCTGAAAAGGAACTTAGTGAAA 1385 Left AAACTTCAGCTTGGAGATAAAATCC 1386 Right CATAGCCTGAAAAGGAACTTAGTGA 1387 Left CTTGGAGATAAAATCCTAGTGATGG 1388 Right TGGTCACTAATTAAGGTTTCCCATA 1389 Left CTTGGAGATAAAATCCTAGTGATGG 1390 Right AGCCTGAAAAGGAACTTAGTGAAAT 1391 Left CTTGGAGATAAAATCCTAGTGATGG 1392 Right TAGCCTGAAAAGGAACTTAGTGAAA 1393 Left GGAACTATTTAAACTTCAGCTTGGA 1394 Right TGGTCACTAATTAAGGTTTCCCATA 1395 Left CTTGGAGATAAAATCCTAGTGATGG 1396 Right CATAGCCTGAAAAGGAACTTAGTGA 1397 Left GGAACTATTTAAACTTCAGCTTGGA 1398 Right AGCCTGAAAAGGAACTTAGTGAAAT 1399 Left GGAACTATTTAAACTTCAGCTTGGA 1400 Right TAGCCTGAAAAGGAACTTAGTGAAA ALK Exon25 401-600 bases 1401 Left TTAATCATTTCCCCTAATCCTTTTC 1402 Right TGGTCACTAATTAAGGTTTCCCATA 1403 Left GAAGCACTTACAACAACACTTAGCA 1404 Right TGGTCACTAATTAAGGTTTCCCATA 1405 Left TGAAGCACTTACAACAACACTTAGC 1406 Right TGGTCACTAATTAAGGTTTCCCATA 1407 Left ATTGTTAAGGCTGTTTCTCTCACAC 1408 Right TGGTCACTAATTAAGGTTTCCCATA 1409 Left GAAGCACTTACAACAACACTTAGCA 1410 Right AGCCTGAAAAGGAACTTAGTGAAAT 1411 Left TGAAGCACTTACAACAACACTTAGC 1412 Right AGCCTGAAAAGGAACTTAGTGAAAT 1413 Left CATTTTTAATCATTTCCCCTAATCC 1414 Right TGGTCACTAATTAAGGTTTCCCATA 1415 Left AGAATTGTTAAGGCTGTTTCTCTCA 1416 Right TGGTCACTAATTAAGGTTTCCCATA 1417 Left ATTGTTAAGGCTGTTTCTCTCACAC 1418 Right AGCCTGAAAAGGAACTTAGTGAAAT 1419 Left GCACCTTGTGTCTTATAAGGTTGTT 1420 Right AGCCTGAAAAGGAACTTAGTGAAAT 1421 Left TTCCATTTCTCTCTTAGTTGTGAGG 1422 Right TGGTCACTAATTAAGGTTTCCCATA 1423 Left GAAGCACTTACAACAACACTTAGCA 1424 Right TAGCCTGAAAAGGAACTTAGTGAAA 1425 Left TGAAGCACTTACAACAACACTTAGC 1426 Right TAGCCTGAAAAGGAACTTAGTGAAA 1427 Left GAAGCACTTACAACAACACTTAGCA 1428 Right CATAGCCTGAAAAGGAACTTAGTGA 1429 Left TGAAGCACTTACAACAACACTTAGC 1430 Right CATAGCCTGAAAAGGAACTTAGTGA 1431 Left ATTGTTAAGGCTGTTTCTCTCACAC 1432 Right TAGCCTGAAAAGGAACTTAGTGAAA 1433 Left GCACCTTGTGTCTTATAAGGTTGTT 1434 Right TAGCCTGAAAAGGAACTTAGTGAAA 1435 Left GAATTGTTAAGGCTGTTTCTCTCAC 1436 Right TGGTCACTAATTAAGGTTTCCCATA 1437 Left ATTGTTAAGGCTGTTTCTCTCACAC 1438 Right CATAGCCTGAAAAGGAACTTAGTGA 1439 Left GCACCTTGTGTCTTATAAGGTTGTT 1440 Right CATAGCCTGAAAAGGAACTTAGTGA ALK Exon25 401-600 bases 1441 Left TTAATCATTTCCCCTAATCCTTTTC 1442 Right TGGTCACTAATTAAGGTTTCCCATA 1443 Left GAAGCACTTACAACAACACTTAGCA 1444 Right TGGTCACTAATTAAGGTTTCCCATA 1445 Left TGAAGCACTTACAACAACACTTAGC 1446 Right TGGTCACTAATTAAGGTTTCCCATA 1447 Left ATTGTTAAGGCTGTTTCTCTCACAC 1448 Right TGGTCACTAATTAAGGTTTCCCATA 1449 Left GAAGCACTTACAACAACACTTAGCA 1450 Right AGCCTGAAAAGGAACTTAGTGAAAT 1451 Left TGAAGCACTTACAACAACACTTAGC 1452 Right AGCCTGAAAAGGAACTTAGTGAAAT 1453 Left CATTTTTAATCATTTCCCCTAATCC 1454 Right TGGTCACTAATTAAGGTTTCCCATA 1455 Left AGAATTGTTAAGGCTGTTTCTCTCA 1456 Right TGGTCACTAATTAAGGTTTCCCATA 1457 Left ATTGTTAAGGCTGTTTCTCTCACAC 1458 Right AGCCTGAAAAGGAACTTAGTGAAAT 1459 Left GCACCTTGTGTCTTATAAGGTTGTT 1460 Right AGCCTGAAAAGGAACTTAGTGAAAT 1461 Left TTCCATTTCTCTCTTAGTTGTGAGG 1462 Right TGGTCACTAATTAAGGTTTCCCATA 1463 Left GAAGCACTTACAACAACACTTAGCA 1464 Right TAGCCTGAAAAGGAACTTAGTGAAA 1465 Left TGAAGCACTTACAACAACACTTAGC 1466 Right TAGCCTGAAAAGGAACTTAGTGAAA 1467 Left GAAGCACTTACAACAACACTTAGCA 1468 Right CATAGCCTGAAAAGGAACTTAGTGA 1469 Left TGAAGCACTTACAACAACACTTAGC 1470 Right CATAGCCTGAAAAGGAACTTAGTGA 1471 Left ATTGTTAAGGCTGTTTCTCTCACAC 1472 Right TAGCCTGAAAAGGAACTTAGTGAAA 1473 Left GCACCTTGTGTCTTATAAGGTTGTT 1474 Right TAGCCTGAAAAGGAACTTAGTGAAA 1475 Left GAATTGTTAAGGCTGTTTCTCTCAC 1476 Right TGGTCACTAATTAAGGTTTCCCATA 1477 Left ATTGTTAAGGCTGTTTCTCTCACAC 1478 Right CATAGCCTGAAAAGGAACTTAGTGA 1479 Left GCACCTTGTGTCTTATAAGGTTGTT 1480 Right CATAGCCTGAAAAGGAACTTAGTGA ALK Exon25 601-800 bases 1481 Left GGACAGTAATAGCACCTTGTGTCTT 1482 Right TGGTCACTAATTAAGGTTTCCCATA 1483 Left AAATGAGGACAGTAATAGCACCTTG 1484 Right TGGTCACTAATTAAGGTTTCCCATA 1485 Left GGACAGTAATAGCACCTTGTGTCTT 1486 Right AGCCTGAAAAGGAACTTAGTGAAAT 1487 Left AAATGAGGACAGTAATAGCACCTTG 1488 Right AGCCTGAAAAGGAACTTAGTGAAAT 1489 Left GCACCTTGTGTCTTATAAGGTTGTT 1490 Right TGGTCACTAATTAAGGTTTCCCATA 1491 Left GGACAGTAATAGCACCTTGTGTCTT 1492 Right TAGCCTGAAAAGGAACTTAGTGAAA 1493 Left AAATGAGGACAGTAATAGCACCTTG 1494 Right TAGCCTGAAAAGGAACTTAGTGAAA 1495 Left AACTTCAGCTTGGAGATAAAATCCT 1496 Right AGTATCCAAGTTATCCCATGTCTCA 1497 Left AACTTCAGCTTGGAGATAAAATCCT 1498 Right GTATCCAAGTTATCCCATGTCTCAG 1499 Left GGACAGTAATAGCACCTTGTGTCTT 1500 Right CATAGCCTGAAAAGGAACTTAGTGA 1501 Left AAATGAGGACAGTAATAGCACCTTG 1502 Right CATAGCCTGAAAAGGAACTTAGTGA 1503 Left TCCTAGGGATAAAATTAGGAAATGC 1504 Right AGTATCCAAGTTATCCCATGTCTCA 1505 Left TCCTAGGGATAAAATTAGGAAATGC 1506 Right GTATCCAAGTTATCCCATGTCTCAG 1507 Left TCCTAGGGATAAAATTAGGAAATGC 1508 Right TCAATAAAGCTCACTTTGAAGGTCT 1509 Left TCCTAGGGATAAAATTAGGAAATGC 1510 Right AGTTGAGGAAGTTCAATAAAGCTCA 1511 Left TCCTAGGGATAAAATTAGGAAATGC 1512 Right TTGAGGAAGTTCAATAAAGCTCACT 1513 Left TCCTAGGGATAAAATTAGGAAATGC 1514 Right TGAGGAAGTTCAATAAAGCTCACTT 1515 Left TCCTAGGGATAAAATTAGGAAATGC 1516 Right CTGAGGAGTTGAGGAAGTTCAATAA 1517 Left TCCTAGGGATAAAATTAGGAAATGC 1518 Right GGAAGTTCAATAAAGCTCACTTTGA 1519 Left TCCTAGGGATAAAATTAGGAAATGC 1520 Right TTCAATAAAGCTCACTTTGAAGGTC ALK Exon25 801-1000 bases 1521 Left AACTTCAGCTTGGAGATAAAATCCT 1522 Right AGTACTGAGGAGTTGAGGAAGTTCA 1523 Left TTAATCATTTCCCCTAATCCTTTTC 1524 Right AGTACTGAGGAGTTGAGGAAGTTCA 1525 Left TTAATCATTTCCCCTAATCCTTTTC 1526 Right TGAGTACTGAGGAGTTGAGGAAGTT 1527 Left AACTTCAGCTTGGAGATAAAATCCT 1528 Right TCAATAAAGCTCACTTTGAAGGTCT 1529 Left AACTTCAGCTTGGAGATAAAATCCT 1530 Right TTGAGGAAGTTCAATAAAGCTCACT 1531 Left AACTTCAGCTTGGAGATAAAATCCT 1532 Right TGAGGAAGTTCAATAAAGCTCACTT 1533 Left TTAATCATTTCCCCTAATCCTTTTC 1534 Right AGTATCCAAGTTATCCCATGTCTCA 1535 Left TTAATCATTTCCCCTAATCCTTTTC 1536 Right GTATCCAAGTTATCCCATGTCTCAG 1537 Left TCCTAGGGATAAAATTAGGAAATGC 1538 Right AGTACTGAGGAGTTGAGGAAGTTCA 1539 Left TCCTAGGGATAAAATTAGGAAATGC 1540 Right TGAGTACTGAGGAGTTGAGGAAGTT 1541 Left TTAATCATTTCCCCTAATCCTTTTC 1542 Right TCAATAAAGCTCACTTTGAAGGTCT 1543 Left TTAATCATTTCCCCTAATCCTTTTC 1544 Right AGTTGAGGAAGTTCAATAAAGCTCA 1545 Left TTAATCATTTCCCCTAATCCTTTTC 1546 Right TGAGGAAGTTCAATAAAGCTCACTT 1547 Left TTAATCATTTCCCCTAATCCTTTTC 1548 Right TTGAGGAAGTTCAATAAAGCTCACT 1549 Left AACTTCAGCTTGGAGATAAAATCCT 1550 Right CTGAGGAGTTGAGGAAGTTCAATAA 1551 Left GGGAAGGAACTATTTAAACTTCAGC 1552 Right AGTATCCAAGTTATCCCATGTCTCA 1553 Left GGGAAGGAACTATTTAAACTTCAGC 1554 Right GTATCCAAGTTATCCCATGTCTCAG 1555 Left AACTTCAGCTTGGAGATAAAATCCT 1556 Right TTCAATAAAGCTCACTTTGAAGGTC 1557 Left AACTTCAGCTTGGAGATAAAATCCT 1558 Right GGAAGTTCAATAAAGCTCACTTTGA 1559 Left AACTTCAGCTTGGAGATAAAATCCT 1560 Right GTCTTTCCACATCAAGTATCCAAGT ALK Exon25 2 kb 1561 Left GGACAGTAATAGCACCTTGTGTCTT 1562 Right GTGCTGAGGACATAAATAGGTCAGT 1563 Left AAAATCATGGACAAAAGAACCATAA 1564 Right TTAAAGCTCCATATAACGATTGCTC 1565 Left GGACAGTAATAGCACCTTGTGTCTT 1566 Right AGTCTCTCTCTCCCAAGGATATTGT 1567 Left AAAATCATGGACAAAAGAACCATAA 1568 Right AGTCTCACTTATTCCCCAAAGAGTT 1569 Left ATCACTTTTTAAAACAACCATTCCA 1570 Right TGGTCACTAATTAAGGTTTCCCATA 1571 Left TCACTTTTTAAAACAACCATTCCAT 1572 Right TGGTCACTAATTAAGGTTTCCCATA 1573 Left TGTAGCTTAGCAAGGGCTTTAGATA 1574 Right TTAAAGCTCCATATAACGATTGCTC 1575 Left AAATGAGGACAGTAATAGCACCTTG 1576 Right TGCATTGCAATATAGAAAACACAGT 1577 Left AAAATCATGGACAAAAGAACCATAA 1578 Right GGTGTCTGGATCAGTCTCACTTATT 1579 Left AAAATCATGGACAAAAGAACCATAA 1580 Right GGAACTAGAGGCTAGGAAGAGAAGA 1581 Left AAATGAGGACAGTAATAGCACCTTG 1582 Right GTGCTGAGGACATAAATAGGTCAGT 1583 Left AACTTCAGCTTGGAGATAAAATCCT 1584 Right TGCATTGCAATATAGAAAACACAGT 1585 Left AAATGAGGACAGTAATAGCACCTTG 1586 Right AGTCTCTCTCTCCCAAGGATATTGT 1587 Left GTCACTCTCCCAACTCTTGATGTAT 1588 Right TGGTCACTAATTAAGGTTTCCCATA 1589 Left AACTTCAGCTTGGAGATAAAATCCT 1590 Right GTGCTGAGGACATAAATAGGTCAGT 1591 Left TGTAGCTTAGCAAGGGCTTTAGATA 1592 Right AGTCTCACTTATTCCCCAAAGAGTT 1593 Left TGTAGCTTAGCAAGGGCTTTAGATA 1594 Right GGTGTCTGGATCAGTCTCACTTATT 1595 Left TGTAGCTTAGCAAGGGCTTTAGATA 1596 Right GGAACTAGAGGCTAGGAAGAGAAGA 1597 Left GGACAGTAATAGCACCTTGTGTCTT 1598 Right TCAGTGACACAAATGAAGAATTGAT 1599 Left TTAATCATTTCCCCTAATCCTTTTC 1600 Right TGCATTGCAATATAGAAAACACAGT ALK Exon24-25 5 kb 1601 Left ATCTCCTTTTGAATGAAAGAGACCT 1602 Right TGGTCACTAATTAAGGTTTCCCATA 1603 Left ATCTCCTTTTGAATGAAAGAGACCT 1604 Right AGCCTGAAAAGGAACTTAGTGAAAT 1605 Left ACGTCAGGGATTTAGGAATTAAAAG 1606 Right TGGTCACTAATTAAGGTTTCCCATA 1607 Left ATGTGAATCATACTCCTCCAGGTAA 1608 Right TGGTCACTAATTAAGGTTTCCCATA 1609 Left ATCTCCTTTTGAATGAAAGAGACCT 1610 Right GTATCCAAGTTATCCCATGTCTCAG 1611 Left ATCTCCTTTTGAATGAAAGAGACCT 1612 Right TAGCCTGAAAAGGAACTTAGTGAAA 1613 Left ACGTCAGGGATTTAGGAATTAAAAG 1614 Right AGCCTGAAAAGGAACTTAGTGAAAT 1615 Left ATCTCCTTTTGAATGAAAGAGACCT 1616 Right CATAGCCTGAAAAGGAACTTAGTGA 1617 Left AGGTATGTGAATCATACTCCTCCAG 1618 Right TGGTCACTAATTAAGGTTTCCCATA 1619 Left AAGAGTCACCAGCTTAAACAAACAC 1620 Right AGCCTGAAAAGGAACTTAGTGAAAT 1621 Left ATGTGAATCATACTCCTCCAGGTAA 1622 Right AGCCTGAAAAGGAACTTAGTGAAAT 1623 Left TGCACAATAAATTAAAAGGGAAAGA 1624 Right TGGTCACTAATTAAGGTTTCCCATA 1625 Left TGTTTATAAATTGGGGGTATTCAAA 1626 Right TGGTCACTAATTAAGGTTTCCCATA 1627 Left TTGTTTATAAATTGGGGGTATTCAA 1628 Right TGGTCACTAATTAAGGTTTCCCATA 1629 Left TGAATCATACTCCTCCAGGTAAATC 1630 Right TGGTCACTAATTAAGGTTTCCCATA 1631 Left AGGTATGTGAATCATACTCCTCCAG 1632 Right AGCCTGAAAAGGAACTTAGTGAAAT 1633 Left ACGTCAGGGATTTAGGAATTAAAAG 1634 Right TAGCCTGAAAAGGAACTTAGTGAAA 1635 Left AAGAGTCACCAGCTTAAACAAACAC 1636 Right TAGCCTGAAAAGGAACTTAGTGAAA 1637 Left ATGTGAATCATACTCCTCCAGGTAA 1638 Right TAGCCTGAAAAGGAACTTAGTGAAA 1639 Left TGCACAATAAATTAAAAGGGAAAGA 1640 Right AGCCTGAAAAGGAACTTAGTGAAAT

TABLE 8 EGFR Capture Primer List for NGS Panel Seq. ID Primer Sequence EGFR Exon18 100-200 bases 1641 Left TGCCAAAGAAGTAGAATGAG 1642 Right AAAGCATCTTCACCCACAGC 1643 Left TGCCAAAGAAGTAGAATGAG 1644 Right TTCTTGACGAGGTCCATGTG 1645 Left TGCCAAAGAAGTAGAATGAG 1646 Right GTCAGAAATGCAGGAAAGCA 1647 Left TGCCAAAGAAGTAGAATGAG 1648 Right AGTCAGAAATGCAGGAAAGCA 1649 Left TGCCAAAGAAGTAGAATGAG 1650 Right CAGTCAGAAATGCAGGAAAGC 1651 Left TGCCAAAGAAGTAGAATGAG 1652 Right ATTCTTGACGAGGTCCATGTG 1653 Left TGCCAAAGAAGTAGAATGAG 1654 Right CATTCTTGACGAGGTCCATGT 1655 Left TGCCAAAGAAGTAGAATGAG 1656 Right GGAAAGCATCTTCACCCACA 1657 Left TGCCAAAGAAGTAGAATGAG 1658 Right CAGCAGTGTGGTCATTCTTGA 1659 Left TGCCAAAGAAGTAGAATGAG 1660 Right AGGACAGTCAGAAATGCAGGA 1661 Left TGCCAAAGAAGTAGAATGAG 1662 Right GGACAGTCAGAAATGCAGGA 1663 Left GCCAAAGAAGTAGAATGAGA 1664 Right AAAGCATCTTCACCCACAGC 1665 Left TGCCAAAGAAGTAGAATGAG 1666 Right GGACAGTCAGAAATGCAGGAA 1667 Left GCCAAAGAAGTAGAATGAGA 1668 Right TTCTTGACGAGGTCCATGTG 1669 Left TGCCAAAGAAGTAGAATGAG 1670 Right TGCAGGAAAGCATCTTCACC 1671 Left TGCCAAAGAAGTAGAATGAG 1672 Right TCAGAAATGCAGGAAAGCATC 1673 Left TGCCAAAGAAGTAGAATGAG 1674 Right GTCATTCTTGACGAGGTCCA 1675 Left TGCCAAAGAAGTAGAATGAG 1676 Right TGGTCATTCTTGACGAGGTC 1677 Left TGCCAAAGAAGTAGAATGAG 1678 Right AGCATCTTCACCCACAGCA 1679 Left TGCCAAAGAAGTAGAATGAG 1680 Right GCATCTTCACCCACAGCAG EGFR Exon18 200-400 bases 1681 Left TGCCAAAGAAGTAGAATGAG 1682 Right CCAGCACTGTGTGTCCAACT 1683 Left TGCCAAAGAAGTAGAATGAG 1684 Right TCCCTCCACTGAGGACAAAG 1685 Left TGCCAAAGAAGTAGAATGAG 1686 Right CTTTCCCTCCACTGAGGACA 1687 Left TGCCAAAGAAGTAGAATGAG 1688 Right CCAACTTTCCCTCCACTGAG 1689 Left TGCCAAAGAAGTAGAATGAG 1690 Right CAAAACCAGTGGAACCAAGG 1691 Left TGCCAAAGAAGTAGAATGAG 1692 Right TGTCCAACTTTCCCTCCACT 1693 Left TGCCAAAGAAGTAGAATGAG 1694 Right GTCCAACTTTCCCTCCACTG 1695 Left TGCCAAAGAAGTAGAATGAG 1696 Right GCAAAACCAGTGGAACCAAG 1697 Left TGCCAAAGAAGTAGAATGAG 1698 Right AGCAAAACCAGTGGAACCAA 1699 Left TGCCAAAGAAGTAGAATGAG 1700 Right AAACCAGTGGAACCAAGGAA 1701 Left TGCCAAAGAAGTAGAATGAG 1702 Right AAAACCAGTGGAACCAAGGA 1703 Left TGCCAAAGAAGTAGAATGAG 1704 Right GTGTCCAACTTTCCCTCCAC 1705 Left TGCCAAAGAAGTAGAATGAG 1706 Right GGCCCAGAGCCATAGAAACT 1707 Left TGCCAAAGAAGTAGAATGAG 1708 Right TTCCCTCCACTGAGGACAAA 1709 Left TGCCAAAGAAGTAGAATGAG 1710 Right TCCAACTTTCCCTCCACTGA 1711 Left TGCCAAAGAAGTAGAATGAG 1712 Right CCCTCCACTGAGGACAAAGT 1713 Left TGCCAAAGAAGTAGAATGAG 1714 Right AACCAGCTGGGCAGTCTCT 1715 Left TGCCAAAGAAGTAGAATGAG 1716 Right GAAACCCTGGCTGAGGGTAG 1717 Left GCCAAAGAAGTAGAATGAGA 1718 Right CCAGCACTGTGTGTCCAACT 1719 Left GCCAAAGAAGTAGAATGAGA 1720 Right TCCCTCCACTGAGGACAAAG EGFR Exon18 400-1000 bases 1721 Left TGCCAAAGAAGTAGAATGAG 1722 Right CAGTGTGGAGTGGGGAAGTT 1723 Left TGCCAAAGAAGTAGAATGAG 1724 Right ACTCCCCTATGCTGGAGGTT 1725 Left TGCCAAAGAAGTAGAATGAG 1726 Right TGGGAAAGAAAGCAAGGAGA 1727 Left TGCCAAAGAAGTAGAATGAG 1728 Right TCTGGGAAAGAAAGCAAGGA 1729 Left TGCCAAAGAAGTAGAATGAG 1730 Right ACCAATGGGGTAAGTGGACA 1731 Left TGCCAAAGAAGTAGAATGAG 1732 Right CCTCGATCATGTGACACTGG 1733 Left TGCCAAAGAAGTAGAATGAG 1734 Right AAAATGGCAAACAGGTGCTC 1735 Left TGCCAAAGAAGTAGAATGAG 1736 Right AACTGGCCAGAGCTGATGTT 1737 Left TGCCAAAGAAGTAGAATGAG 1738 Right AAACTGGCCAGAGCTGATGT 1739 Left TGCCAAAGAAGTAGAATGAG 1740 Right ACGCCATCGAGAGTAACACC 1741 Left TGCCAAAGAAGTAGAATGAG 1742 Right AGGAGCATGCCAAAATGAAG 1743 Left TGCCAAAGAAGTAGAATGAG 1744 Right TGTTGAAGGAAGCCCTTTTG 1745 Left TGCCAAAGAAGTAGAATGAG 1746 Right CCAATGGGGTAAGTGGACAG 1747 Left TGCCAAAGAAGTAGAATGAG 1748 Right TTGCCTTCTTCCTCGATCAT 1749 Left TGCCAAAGAAGTAGAATGAG 1750 Right CATCGAACAGAAAGGCCACT 1751 Left TGCCAAAGAAGTAGAATGAG 1752 Right GGTGGCAGGAGAGAGAGCTA 1753 Left TGCCAAAGAAGTAGAATGAG 1754 Right ATGGGACCAATGGGGTAAGT 1755 Left TGCCAAAGAAGTAGAATGAG 1756 Right TGGAGGTTGTCATCGAACAG 1757 Left TGCCAAAGAAGTAGAATGAG 1758 Right CTGGAGGTTGTCATCGAACA 1759 Left TGCCAAAGAAGTAGAATGAG 1760 Right TATGCTGGAGGTTGTCATCG EGFR Exon19 100-200 bases 1761 Left CTTCCTTGTTCCTCCACCTCAT 1762 Right ACCCAGGACTGGCACTCAC 1763 Left CTTCCTTGTTCCTCCACCTCAT 1764 Right CCCAGGACTGGCACTCAC 1765 Left CTTCCTTGTTCCTCCACCTCAT 1766 Right ACCCAGGACTGGCACTCA 1767 Left CTTGTTCCTCCACCTCATTCC 1768 Right ACCCAGGACTGGCACTCAC 1769 Left CCTTGTTCCTCCACCTCATTC 1770 Right ACCCAGGACTGGCACTCAC 1771 Left TCCTTGTTCCTCCACCTCATT 1772 Right ACCCAGGACTGGCACTCAC 1773 Left TTCCTTGTTCCTCCACCTCAT 1774 Right ACCCAGGACTGGCACTCAC 1775 Left CTTCCTTGTTCCTCCACCTCATT 1776 Right ACCCAGGACTGGCACTCAC 1777 Left CAACCTCACCCTTCCTTGTTC 1778 Right ACCCAGGACTGGCACTCAC 1779 Left CTTGTTCCTCCACCTCATTCC 1780 Right CCCAGGACTGGCACTCAC 1781 Left CCTTGTTCCTCCACCTCATTC 1782 Right CCCAGGACTGGCACTCAC 1783 Left CCTTGTTCCTCCACCTCATTC 1784 Right ACCCAGGACTGGCACTCA 1785 Left CTTGTTCCTCCACCTCATTCC 1786 Right ACCCAGGACTGGCACTCA 1787 Left TCCTTGTTCCTCCACCTCATT 1788 Right CCCAGGACTGGCACTCAC 1789 Left TCCTTGTTCCTCCACCTCATT 1790 Right ACCCAGGACTGGCACTCA 1791 Left TTCCTTGTTCCTCCACCTCAT 1792 Right CCCAGGACTGGCACTCAC 1793 Left TTCCTTGTTCCTCCACCTCAT 1794 Right ACCCAGGACTGGCACTCA 1795 Left CTTCCTTGTTCCTCCACCTCATT 1796 Right CCCAGGACTGGCACTCAC 1797 Left CTTCCTTGTTCCTCCACCTCATT 1798 Right ACCCAGGACTGGCACTCA 1799 Left CAACCTCACCCTTCCTTGTTC 1800 Right CCCAGGACTGGCACTCAC EGFR Exon19 200-400 bases 1801 Left AAGATCATTCTACAAGATGTCAGTGG 1802 Right AACTGCACATTCAGAGATTCTTTCT 1803 Left AGATCATTCTACAAGATGTCAGTGC 1804 Right AACTGCACATTCAGAGATTCTTTCT 1805 Left TCCAAGATCATTCTACAAGATGTCA 1806 Right ACATTCAGAGATTCTTTCTGCATCA 1807 Left TCCAAGATCATTCTACAAGATGTCA 1808 Right ATTCAGAGATTCTTTCTGCATCATA 1809 Left TCCAAGATCATTCTACAAGATGTCA 1810 Right CATTCAGAGATTCTTTCTGCATCA 1811 Left CAAGATCATTCTACAAGATGTCAGTG 1812 Right AACTGCACATTCAGAGATTCTTTCT 1813 Left TCCAAGATCATTCTACAAGATGTCA 1814 Right CATTCAGAGATTCTTTCTGCATCATA 1815 Left AAGATCATTCTACAAGATGTCAGTGG 1816 Right CATTCAGAGATTCTTTCTGCATCAT 1817 Left AAGATCATTCTACAAGATGTCAGTGG 1818 Right TAACTGCACATTCAGAGATTCTTTC 1819 Left AGATCATTCTACAAGATGTCAGTGC 1820 Right CATTCAGAGATTCTTTCTGCATCAT 1821 Left AGATCATTCTACAAGATGTCAGTGC 1822 Right TAACTGCACATTCAGAGATTCTTTC 1823 Left AAGATCATTCTACAAGATGTCAGTGC 1824 Right TAACTGCACATTCAGAGATTCTTTCT 1825 Left AGATCATTCTACAAGATGTCAGTGC 1826 Right TAACTGCACATTCAGAGATTCTTTCT 1827 Left TCCAAGATCATTCTACAAGATGTCA 1828 Right TTCAGAGATTCTTTCTGCATCATAATA 1829 Left TCCAAGATCATTCTACAAGATGTCA 1830 Right ACTGCACATTCAGAGATTCTTTCT 1831 Left TCCAAGATCATTCTACAAGATGTCA 1832 Right GCACATTCAGAGATTCTTTCTGC 1833 Left TCCAAGATCATTCTACAAGATGTCA 1834 Right TCAGAGATTCTTTCTGCATCATAATA 1835 Left CCAAGATCATTCTACAAGATGTCAGT 1836 Right ACATTCAGAGATTCTTTCTGCATCA 1837 Left CCAAGATCATTCTACAAGATGTCAGT 1838 Right ATTCAGAGATTCTTTCTGCATCATA 1839 Left AAGATCATTCTACAAGATGTCAGTGC 1840 Right ACATTCAGAGATTCTTTCTGCATCA EGFR Exon19 400-1000 bases 1841 Left AATACCAATCCATGAAAAAGCATTA 1842 Right AACATGTCACCAACTGGGTATAACT 1843 Left CCTATTCCTTTATAACCCCTTTCAA 1844 Right AACATGTCACCAACTGGGTATAACT 1845 Left TCCAAGATCATTCTACAAGATGTCA 1846 Right AACATGTCACCAACTGGGTATAACT 1847 Left TTTCAAGCTCGTTCAGAGAGTATTT 1848 Right AACATGTCACCAACTGGGTATAACT 1849 Left TTCAGAGAGTATTTCACACAATCCA 1850 Right AACATGTCACCAACTGGGTATAACT 1851 Left GTGTCTCACTTTCCAAGATCATTCT 1852 Right AACATGTCACCAACTGGGTATAACT 1853 Left AGTGTCTCACTTTCCAAGATCATTC 1854 Right AACATGTCACCAACTGGGTATAACT 1855 Left CCATGAAAAAGCATTATTGAAGTCT 1856 Right AACATGTCACCAACTGGGTATAACT 1857 Left TATTCCTTTATAACCCCTTTCAAGC 1858 Right AACATGTCACCAACTGGGTATAACT 1859 Left ATGGAAATACTCTTGGAATGAACAA 1860 Right AACATGTCACCAACTGGGTATAACT 1861 Left CTCGTTCAGAGAGTATTTCACACAA 1862 Right AACATGTCACCAACTGGGTATAACT 1863 Left TCCAAGATCATTCTACAAGATGTCA 1864 Right ACTGAACAGCTACCTTTCAACAAAC 1865 Left CCTATTCCTTTATAACCCCTTTCAA 1866 Right AACTGCACATTCAGAGATTCTTTCT 1867 Left TCCAAGATCATTCTACAAGATGTCA 1868 Right AACTGCACATTCAGAGATTCTTTCT 1869 Left ACTCTTGGAATGAACAAAATACCAA 1870 Right AACATGTCACCAACTGGGTATAACT 1871 Left CCTATTCCTTTATAACCCCTTTCAA 1872 Right GGGTATAACTGCACATTCAGAGATT 1873 Left TCCAAGATCATTCTACAAGATGTCA 1874 Right GGGTATAACTGCACATTCAGAGATT 1875 Left AGTGTCTCACTTTCCAAGATCATTC 1876 Right ACTGAACAGCTACCTTTCAACAAAC 1877 Left GTGTCTCACTTTCCAAGATCATTCT 1878 Right ACTGAACAGCTACCTTTCAACAAAC 1879 Left AGTGTCTCACTTTCCAAGATCATTC 1880 Right AACTGCACATTCAGAGATTCTTTCT EGFR Exon20 100-200 bases 1881 Left GTGACCCTTGTCTCTGTGTTCTT 1882 Right CCTGTGCCAGGGACCTTAC 1883 Left ACCCTTGTCTCTGTGTTCTTGTC 1884 Right CCTGTGCCAGGGACCTTAC 1885 Left GACCCTTGTCTCTGTGTTCTTGT 1886 Right CCTGTGCCAGGGACCTTAC 1887 Left GACCCTTGTCTCTGTGTTCTTGTC 1888 Right CCTGTGCCAGGGACCTTAC 1889 Left GTGACCCTTGTCTCTGTGTTCTTGT 1890 Right CCTGTGCCAGGGACCTTAC 1891 Left AGGTGACCCTTGTCTCTGTGTT 1892 Right CCTGTGCCAGGGACCTTAC 1893 Left GACCCTTGTCTCTGTGTTCTTGT 1894 Right CCTGTGCCAGGGACCTTA 1895 Left GTGACCCTTGTCTCTGTGTTCTT 1896 Right CCTGTGCCAGGGACCTTA 1897 Left ACCCTTGTCTCTGTGTTCTTGTC 1898 Right CCTGTGCCAGGGACCTTA 1899 Left GACCCTTGTCTCTGTGTTCTTGTC 1900 Right CCTGTGCCAGGGACCTTA 1901 Left GTGACCCTTGTCTCTGTGTTCTTG 1902 Right CCTGTGCCAGGGACCTTAC 1903 Left TGACCCTTGTCTCTGTGTTCTTGT 1904 Right CCTGTGCCAGGGACCTTAC 1905 Left GAGGTGACCCTTGTCTCTGTGT 1906 Right CCTGTGCCAGGGACCTTAC 1907 Left TGACCCTTGTCTCTGTGTTCTTG 1908 Right CCTGTGCCAGGGACCTTAC 1909 Left AGGTGACCCTTGTCTCTGTGTTCT 1910 Right CCTGTGCCAGGGACCTTAC 1911 Left CTGAGGTGACCCTTGTCTCTGT 1912 Right CCTGTGCCAGGGACCTTAC 1913 Left AGGTGACCCTTGTCTCTGTGTTCTT 1914 Right CCTGTGCCAGGGACCTTAC 1915 Left GGTGACCCTTGTCTCTGTGTTCT 1916 Right CCTGTGCCAGGGACCTTAC 1917 Left AGGTGACCCTTGTCTCTGTGTTC 1918 Right CCTGTGCCAGGGACCTTAC 1919 Left GAGGTGACCCTTGTCTCTGTGTT 1920 Right CCTGTGCCAGGGACCTTAC EGFR Exon20 200-400 bases 1921 Left AAGCTCTGTAGAGAAGGCGTACAT 1922 Right AAATATACAGCTTGCAAGGACTCTG 1923 Left AGCTCTGTAGAGAAGGCGTACATT 1924 Right AAATATACAGCTTGCAAGGACTCTG 1925 Left TCTGTAGAGAAGGCGTACATTTGTC 1926 Right AAATATACAGCTTGCAAGGACTCTG 1927 Left GTAGAGAAGGCGTACATTTGTCCT 1928 Right AAATATACAGCTTGCAAGGACTCTG 1929 Left AAGCTCTGTAGAGAAGGCGTACATT 1930 Right AAATATACAGCTTGCAAGGACTCTG 1931 Left GCTCTGTAGAGAAGGCGTACATTT 1932 Right AAATATACAGCTTGCAAGGACTCTG 1933 Left CTCTGTAGAGAAGGCGTACATTTG 1934 Right AAATATACAGCTTGCAAGGACTCTG 1935 Left CTGTAGAGAAGGCGTACATTTGTC 1936 Right AAATATACAGCTTGCAAGGACTCTG 1937 Left TTCTGTCAAGCTCTGTAGAGAAGG 1938 Right AAATATACAGCTTGCAAGGACTCTG 1939 Left AAGCTCTGTAGAGAAGGCGTACA 1940 Right AAATATACAGCTTGCAAGGACTCTG 1941 Left TACATTTGTCCTTCCAAATGAGC 1942 Right AAATATACAGCTTGCAAGGACTCTG 1943 Left CAAGCTCTGTAGAGAAGGCGTACAT 1944 Right AAATATACAGCTTGCAAGGACTCTG 1945 Left AGCTCTGTAGAGAAGGCGTACATT 1946 Right GGAAATATACAGCTTGCAAGGACTC 1947 Left AAGCTCTGTAGAGAAGGCGTACAT 1948 Right GGAAATATACAGCTTGCAAGGACTC 1949 Left GTCAAGCTCTGTAGAGAAGGCGTA 1950 Right AAATATACAGCTTGCAAGGACTCTG 1951 Left TCTGTAGAGAAGGCGTACATTTGTC 1952 Right GGAAATATACAGCTTGCAAGGACTC 1953 Left TGTAGAGAAGGCGTACATTTGTCCT 1954 Right AAATATACAGCTTGCAAGGACTCTG 1955 Left GTAGAGAAGGCGTACATTTGTCCT 1956 Right GGAAATATACAGCTTGCAAGGACTC 1957 Left AAGCTCTGTAGAGAAGGCGTACATT 1958 Right GGAAATATACAGCTTGCAAGGACTC 1959 Left AGCTCTGTAGAGAAGGCGTACATT 1960 Right ATGGAAATATACAGCTTGCAAGGAC EGFR Exon20 400-1000 bases 1961 Left TTTCTACCAACTTCTGTCAAGCTCT 1962 Right ATCTAGAAGAAGCAAACGAAGATGA 1963 Left GTTTCTACCAACTTCTGTCAAGCTC 1964 Right ATCTAGAAGAAGCAAACGAAGATGA 1965 Left GTTTCTACCAACTTCTGTCAAGCTC 1966 Right GATCTAGAAGAAGCAAACGAAGATG 1967 Left GTTTCTACCAACTTCTGTCAAGCTC 1968 Right CTATGACAGAGAGAGAAGGAAGACG 1969 Left CTGTGTTTCTACCAACTTCTGTCAA 1970 Right GATCTAGAAGAAGCAAACGAAGATG 1971 Left TGTGTTTCTACCAACTTCTGTCAAG 1972 Right ATCTAGAAGAAGCAAACGAAGATGA 1973 Left TGTGTTTCTACCAACTTCTGTCAAG 1974 Right GATCTAGAAGAAGCAAACGAAGATG 1975 Left CTGTGTTTCTACCAACTTCTGTCAA 1976 Right ATCTAGAAGAAGCAAACGAAGATGA 1977 Left AGAAAGAATCTCTGAATGTGCAGTT 1978 Right AAATATACAGCTTGCAAGGACTCTG 1979 Left GAAATTGTGTTTGTTGAAAGGTAGC 1980 Right ATCTAGAAGAAGCAAACGAAGATGA 1981 Left GAAATTGTGTTTGTTGAAAGGTAGC 1982 Right GATCTAGAAGAAGCAAACGAAGATG 1983 Left AATCTCTGAATGTGCAGTTATACCC 1984 Right AAATATACAGCTTGCAAGGACTCTG 1985 Left TTTCTACCAACTTCTGTCAAGCTCT 1986 Right GTTATAAAGTCCGTGTGGATCATTT 1987 Left CTGTGTTTCTACCAACTTCTGTCAA 1988 Right AAATATACAGCTTGCAAGGACTCTG 1989 Left TTTCTACCAACTTCTGTCAAGCTCT 1990 Right TTATAAAGTCCGTGTGGATCATTTC 1991 Left GAAATTGTGTTTGTTGAAAGGTAGC 1992 Right AAATATACAGCTTGCAAGGACTCTG 1993 Left TTTCTACCAACTTCTGTCAAGCTCT 1994 Right CTGTTATAAAGTCCGTGTGGATCAT 1995 Left GTTTCTACCAACTTCTGTCAAGCTC 1996 Right GTTATAAAGTCCGTGTGGATCATTT 1997 Left TTTCTACCAACTTCTGTCAAGCTCT 1998 Right TCTAGAAGAAGCAAACGAAGATGAG 1999 Left TTTCTACCAACTTCTGTCAAGCTCT 2000 Right CTCCACGAATCACACTGATTATTTA EGFR Exon21 100-200 bases 2001 Left ACGTCTTCCTTCTCTCTCTGTCATA 2002 Right ACACAGCAAAGCAGAAACTCAC 2003 Left TTAACGTCTTCCTTCTCTCTCTGTC 2004 Right ACACAGCAAAGCAGAAACTCAC 2005 Left TAACGTCTTCCTTCTCTCTCTGTCA 2006 Right ACACAGCAAAGCAGAAACTCAC 2007 Left ACGTCTTCCTTCTCTCTCTGTCATA 2008 Right CACACAGCAAAGCAGAAACTCAC 2009 Left ACGTCTTCCTTCTCTCTCTGTCAT 2010 Right ACACAGCAAAGCAGAAACTCAC 2011 Left CCAGTTAACGTCTTCCTTCTCTCTC 2012 Right ACACAGCAAAGCAGAAACTCAC 2013 Left ACGTCTTCCTTCTCTCTCTGTCATA 2014 Right CCACACAGCAAAGCAGAAACT 2015 Left AACGTCTTCCTTCTCTCTCTGTCAT 2016 Right ACACAGCAAAGCAGAAACTCAC 2017 Left CAGTTAACGTCTTCCTTCTCTCTCT 2018 Right ACACAGCAAAGCAGAAACTCAC 2019 Left AGTTAACGTCTTCCTTCTCTCTCTG 2020 Right ACACAGCAAAGCAGAAACTCAC 2021 Left ACGTCTTCCTTCTCTCTCTGTCATA 2022 Right CACACAGCAAAGCAGAAACTCA 2023 Left GTTAACGTCTTCCTTCTCTCTCTGT 2024 Right ACACAGCAAAGCAGAAACTCAC 2025 Left TTAACGTCTTCCTTCTCTCTCTGTC 2026 Right CACACAGCAAAGCAGAAACTCAC 2027 Left GTTAACGTCTTCCTTCTCTCTCTGTC 2028 Right ACACAGCAAAGCAGAAACTCAC 2029 Left AGTTAACGTCTTCCTTCTCTCTCTGT 2030 Right ACACAGCAAAGCAGAAACTCAC 2031 Left TTAACGTCTTCCTTCTCTCTCTGTC 2032 Right CCACACAGCAAAGCAGAAACT 2033 Left CCAGTTAACGTCTTCCTTCTCTCT 2034 Right ACACAGCAAAGCAGAAACTCAC 2035 Left TTAACGTCTTCCTTCTCTCTCTGTC 2036 Right CACACAGCAAAGCAGAAACTCA 2037 Left TAACGTCTTCCTTCTCTCTCTGTCA 2038 Right CACACAGCAAAGCAGAAACTCAC 2039 Left CGTCTTCCTTCTCTCTCTGTCATA 2040 Right ACACAGCAAAGCAGAAACTCAC EGFR Exon21 200-400 bases 2041 Left ACGTCTTCCTTCTCTCTCTGTCATA 2042 Right TGTCTCTAAGGGGAGGGAGTTATAC 2043 Left ACGTCTTCCTTCTCTCTCTGTCATA 2044 Right GAAAGTGAACATTTAGGATGTGGAG 2045 Left ACGTCTTCCTTCTCTCTCTGTCATA 2046 Right AAAGTGAACATTTAGGATGTGGAGA 2047 Left ACGTCTTCCTTCTCTCTCTGTCATA 2048 Right AGAAAGTGAACATTTAGGATGTGGA 2049 Left TTAACGTCTTCCTTCTCTCTCTGTC 2050 Right TGTCTCTAAGGGGAGGGAGTTATAC 2051 Left ACGTCTTCCTTCTCTCTCTGTCATA 2052 Right GTGTCAAGAAACTAGTGCTGGGTAG 2053 Left TTAACGTCTTCCTTCTCTCTCTGTC 2054 Right AAAGTGAACATTTAGGATGTGGAGA 2055 Left TTAACGTCTTCCTTCTCTCTCTGTC 2056 Right GAAAGTGAACATTTAGGATGTGGAG 2057 Left TTAACGTCTTCCTTCTCTCTCTGTC 2058 Right AGAAAGTGAACATTTAGGATGTGGA 2059 Left ACGTCTTCCTTCTCTCTCTGTCATA 2060 Right AGTGAACATTTAGGATGTGGAGATG 2061 Left ACGTCTTCCTTCTCTCTCTGTCATA 2062 Right AAGTGAACATTTAGGATGTGGAGAT 2063 Left ACGTCTTCCTTCTCTCTCTGTCATA 2064 Right GTCAAGAAACTAGTGCTGGGTAGAT 2065 Left TTAACGTCTTCCTTCTCTCTCTGTC 2066 Right GTGTCAAGAAACTAGTGCTGGGTAG 2067 Left TTAACGTCTTCCTTCTCTCTCTGTC 2068 Right AGTGAACATTTAGGATGTGGAGATG 2069 Left TTAACGTCTTCCTTCTCTCTCTGTC 2070 Right AAGTGAACATTTAGGATGTGGAGAT 2071 Left ACGTCTTCCTTCTCTCTCTGTCATA 2072 Right TCAAGAAACTAGTGCTGGGTAGATG 2073 Left TTAACGTCTTCCTTCTCTCTCTGTC 2074 Right GTCAAGAAACTAGTGCTGGGTAGAT 2075 Left ACGTCTTCCTTCTCTCTCTGTCATA 2076 Right GAAAGGGAAAGACATAGAAAGTGAAC 2077 Left ACGTCTTCCTTCTCTCTCTGTCATA 2078 Right TGTCAAGAAACTAGTGCTGGGTAG 2079 Left TAACGTCTTCCTTCTCTCTCTGTCA 2080 Right TGTCTCTAAGGGGAGGGAGTTATAC EGFR Exon21 400-1000 bases 2081 Left ACGTCTTCCTTCTCTCTCTGTCATA 2082 Right CAAAGTAACAATCAACAGACACTGG 2083 Left ACGTCTTCCTTCTCTCTCTGTCATA 2084 Right AAAGATGAGATAACTTGGTGGAGTG 2085 Left ACGTCTTCCTTCTCTCTCTGTCATA 2086 Right CAAAGATGAGATAACTTGGTGGAGT 2087 Left ACGTCTTCCTTCTCTCTCTGTCATA 2088 Right TGAGGTAATAAGTCAGCCATTTTTC 2089 Left ACGTCTTCCTTCTCTCTCTGTCATA 2090 Right CCACAAAGTAACAATCAACAGACAC 2091 Left TTCTAGATCCTCTTTGCATGAAATC 2092 Right AAAGATGAGATAACTTGGTGGAGTG 2093 Left TTCTAGATCCTCTTTGCATGAAATC 2094 Right CAAAGATGAGATAACTTGGTGGAGT 2095 Left ACGTCTTCCTTCTCTCTCTGTCATA 2096 Right GGTAATAAGTCAGCCATTTTTCCTT 2097 Left ACGTCTTCCTTCTCTCTCTGTCATA 2098 Right AATTTCTTTATGCCTCCATTTCTTC 2099 Left ACGTCTTCCTTCTCTCTCTGTCATA 2100 Right GATGAGATAACTTGGTGGAGTGAAT 2101 Left ACGTCTTCCTTCTCTCTCTGTCATA 2102 Right GAATTTTCCAAGAACTTATTCCACA 2103 Left ACGTCTTCCTTCTCTCTCTGTCATA 2104 Right TGTGGAATTTTCCAAGAACTTATTC 2105 Left ACGTCTTCCTTCTCTCTCTGTCATA 2106 Right AATAAGTCAGCCATTTTTCCTTTTC 2107 Left ACGTCTTCCTTCTCTCTCTGTCATA 2108 Right CCATTTCAAAGATGAGATAACTTGG 2109 Left AGGCTTTACAAGCTTGAGATTCTTT 2110 Right CAAAGTAACAATCAACAGACACTGG 2111 Left TTCTAGATCCTCTTTGCATGAAATC 2112 Right AATTTCTTTATGCCTCCATTTCTTC 2113 Left ACGTCTTCCTTCTCTCTCTGTCATA 2114 Right AGATAACTTGGTGGAGTGAATTGAA 2115 Left TTCTAGATCCTCTTTGCATGAAATC 2116 Right GATGAGATAACTTGGTGGAGTGAAT 2117 Left TTCTAGATCCTCTTTGCATGAAATC 2118 Right CCATTTCAAAGATGAGATAACTTGG 2119 Left AGGCTTTACAAGCTTGAGATTCTTT 2120 Right AAAGATGAGATAACTTGGTGGAGTG EGFR Exon22 100-200 bases 2121 Left CACACTGACGTGCCTCTCC 2122 Right CGTATCTCCCTTCCCTGATTAC 2123 Left CCACACTGACGTGCCTCTC 2124 Right CGTATCTCCCTTCCCTGATTAC 2125 Left CACACTGACGTGCCTCTCC 2126 Right CCGTATCTCCCTTCCCTGATTAC 2127 Left CCACACTGACGTGCCTCTC 2128 Right CCGTATCTCCCTTCCCTGATTAC 2129 Left CACACTGACGTGCCTCTCC 2130 Right CCGTATCTCCCTTCCCTGATTA 2131 Left CCACACTGACGTGCCTCTC 2132 Right CCGTATCTCCCTTCCCTGATTA 2133 Left CGAAGCCACACTGACGTG 2134 Right CGTATCTCCCTTCCCTGATTAC 2135 Left CGAAGCCACACTGACGTG 2136 Right CCGTATCTCCCTTCCCTGATTAC 2137 Left ACCATGCGAAGCCACACT 2138 Right CGTATCTCCCTTCCCTGATTAC 2139 Left ACCATGCGAAGCCACACT 2140 Right CCGTATCTCCCTTCCCTGATTAC 2141 Left CGAAGCCACACTGACGTG 2142 Right CCGTATCTCCCTTCCCTGATTA 2143 Left CATGCGAAGCCACACTGAC 2144 Right CGTATCTCCCTTCCCTGATTAC 2145 Left CATGCGAAGCCACACTGA 2146 Right CGTATCTCCCTTCCCTGATTAC 2147 Left CATGCGAAGCCACACTGAC 2148 Right CCGTATCTCCCTTCCCTGATTAC 2149 Left CACACTGACGTGCCTCTCC 2150 Right TATCTCCCCTCCCCGTATCT 2151 Left CATGCGAAGCCACACTGA 2152 Right CCGTATCTCCCTTCCCTGATTAC 2153 Left CACACTGACGTGCCTCTCC 2154 Right CCGTATCTCCCTTCCCTGAT 2155 Left CCACACTGACGTGCCTCTC 2156 Right CCGTATCTCCCTTCCCTGAT 2157 Left ACCATGCGAAGCCACACT 2158 Right CCGTATCTCCCTTCCCTGATTA 2159 Left CACACTGACGTGCCTCTCC 2160 Right TATCTCCCCTCCCCGTATCTC EGFR Exon22 200-400 bases 2161 Left GTATTTTGAAACTCAAGATCGCATT 2162 Right CGTATCTCCCTTCCCTGATTAC 2163 Left CCTCCATGAGTACGTATTTTGAAAC 2164 Right CGTATCTCCCTTCCCTGATTAC 2165 Left GTATTTTGAAACTCAAGATCGCATT 2166 Right CCGTATCTCCCTTCCCTGATTAC 2167 Left CCTCCATGAGTACGTATTTTGAAAC 2168 Right CCGTATCTCCCTTCCCTGATTAC 2169 Left GTATTTTGAAACTCAAGATCGCATT 2170 Right CATGGCAAACTCTTGCTATCC 2171 Left GTATTTTGAAACTCAAGATCGCATT 2172 Right ATATCCCCATGGCAAACTCTT 2173 Left CCATGAGTACGTATTTTGAAACTCA 2174 Right CGTATCTCCCTTCCCTGATTAC 2175 Left GTATTTTGAAACTCAAGATCGCATT 2176 Right CCGTATCTCCCTTCCCTGATTA 2177 Left CCTCCATGAGTACGTATTTTGAAAC 2178 Right CCGTATCTCCCTTCCCTGATTA 2179 Left CCATGAGTACGTATTTTGAAACTCA 2180 Right CCGTATCTCCCTTCCCTGATTAC 2181 Left CCATGAGTACGTATTTTGAAACTCA 2182 Right CATGGCAAACTCTTGCTATCC 2183 Left TATTTTGAAACTCAAGATCGCATTC 2184 Right CGTATCTCCCTTCCCTGATTAC 2185 Left GTATTTTGAAACTCAAGATCGCATT 2186 Right CTTATCTCCCCTCCCCGTATCT 2187 Left GTATTTTGAAACTCAAGATCGCATT 2188 Right ACATATCCCCATGGCAAACTCT 2189 Left CTTTTCCTCCATGAGTACGTATTTT 2190 Right CGTATCTCCCTTCCCTGATTAC 2191 Left TATTTTGAAACTCAAGATCGCATTC 2192 Right CCGTATCTCCCTTCCCTGATTAC 2193 Left GTATTTTGAAACTCAAGATCGCATT 2194 Right ACATATCCCCATGGCAAACTCTT 2195 Left CCTCCATGAGTACGTATTTTGAAAC 2196 Right CTTATCTCCCCTCCCCGTATCT 2197 Left CCATGAGTACGTATTTTGAAACTCA 2198 Right CCGTATCTCCCTTCCCTGATTA 2199 Left GTATTTTGAAACTCAAGATCGCATT 2200 Right ACATATCCCCATGGCAAACTC EGFR Exon22 400-1000 bases 2201 Left GTATTTTGAAACTCAAGATCGCATT 2202 Right TTGAATCCAAAATAAAGGAATGTGT 2203 Left GTATTTTGAAACTCAAGATCGCATT 2204 Right CACACTGAGCACTCAATAAAGAGAA 2205 Left GTATTTTGAAACTCAAGATCGCATT 2206 Right TTCTCCACTACAAATCACCACAGTA 2207 Left GTATTTTGAAACTCAAGATCGCATT 2208 Right ATTCTTCAAAGGTAGCTGATTGATG 2209 Left GTATTTTGAAACTCAAGATCGCATT 2210 Right ATCCAAAATAAAGGAATGTGTGTGT 2211 Left GTATTTTGAAACTCAAGATCGCATT 2212 Right GCTTACCTTGTTATCAAGTCCTGAA 2213 Left TGGTCTATTGAAAGAGCTTATCCAG 2214 Right TTGAATCCAAAATAAAGGAATGTGT 2215 Left CCTCCATGAGTACGTATTTTGAAAC 2216 Right TTGAATCCAAAATAAAGGAATGTGT 2217 Left CCTCCATGAGTACGTATTTTGAAAC 2218 Right CACACTGAGCACTCAATAAAGAGAA 2219 Left CCTCCATGAGTACGTATTTTGAAAC 2220 Right TTCTCCACTACAAATCACCACAGTA 2221 Left TGGTCTATTGAAAGAGCTTATCCAG 2222 Right ATCCAAAATAAAGGAATGTGTGTGT 2223 Left CCTCCATGAGTACGTATTTTGAAAC 2224 Right ATCCAAAATAAAGGAATGTGTGTGT 2225 Left CCTCCATGAGTACGTATTTTGAAAC 2226 Right GCTTACCTTGTTATCAAGTCCTGAA 2227 Left GTATTTTGAAACTCAAGATCGCATT 2228 Right CAAAGGTAGCTGATTGATGAGAGTT 2229 Left GTATTTTGAAACTCAAGATCGCATT 2230 Right TATTCCTTCTCCACTACAAATCACC 2231 Left GTATTTTGAAACTCAAGATCGCATT 2232 Right CCACTACAAATCACCACAGTATTCA 2233 Left GTATTTTGAAACTCAAGATCGCATT 2234 Right CTTGATTGAATCCAAAATAAAGGAA 2235 Left GTATTTTGAAACTCAAGATCGCATT 2236 Right TAAGAACAGAGACATCAGACCACAC 2237 Left CCTCCATGAGTACGTATTTTGAAAC 2238 Right CAAAGGTAGCTGATTGATGAGAGTT 2239 Left GTATTTTGAAACTCAAGATCGCATT 2240 Right AAATTCTTCAAAGGTAGCTGATTGA EGFR Exon18-19 2 kb 2241 Left ATGCCAAAGAAGTAGAATGAGAAAA 2242 Right AACATGTCACCAACTGGGTATAACT 2243 Left ATGCCAAAGAAGTAGAATGAGAAAA 2244 Right GGGTATAACTGCACATTCAGAGATT 2245 Left TCTCCAAAATATATGCCAAAGAAGT 2246 Right AACATGTCACCAACTGGGTATAACT 2247 Left TGCCAAAGAAGTAGAATGAGAAAAA 2248 Right AACATGTCACCAACTGGGTATAACT 2249 Left AATCTCCAAAATATATGCCAAAGAA 2250 Right AACATGTCACCAACTGGGTATAACT 2251 Left AAATCTCCAAAATATATGCCAAAGA 2252 Right AACATGTCACCAACTGGGTATAACT 2253 Left TCTCCAAAATATATGCCAAAGAAGT 2254 Right AACTGCACATTCAGAGATTCTTTCT 2255 Left TCTCCAAAATATATGCCAAAGAAGT 2256 Right GGGTATAACTGCACATTCAGAGATT 2257 Left TGCCAAAGAAGTAGAATGAGAAAAA 2258 Right GGGTATAACTGCACATTCAGAGATT 2259 Left AAATCTCCAAAATATATGCCAAAGA 2260 Right AACTGCACATTCAGAGATTCTTTCT 2261 Left AATCTCCAAAATATATGCCAAAGAA 2262 Right GGGTATAACTGCACATTCAGAGATT 2263 Left AAATCTCCAAAATATATGCCAAAGA 2264 Right GGGTATAACTGCACATTCAGAGATT 2265 Left ATCTCCAAAATATATGCCAAAGAAG 2266 Right AACATGTCACCAACTGGGTATAACT 2267 Left AAAAATCTCCAAAATATATGCCAAAG 2268 Right AACATGTCACCAACTGGGTATAACT 2269 Left AAAATCTCCAAAATATATGCCAAAG 2270 Right AACATGTCACCAACTGGGTATAACT 2271 Left ATGCCAAAGAAGTAGAATGAGAAAA 2272 Right ACTGGGTATAACTGCACATTCAGAG 2273 Left ATCTCCAAAATATATGCCAAAGAAG 2274 Right AACTGCACATTCAGAGATTCTTTCT 2275 Left AATCTCCAAAATATATGCCAAAGAAG 2276 Right AACATGTCACCAACTGGGTATAACT 2277 Left ATCTCCAAAATATATGCCAAAGAAG 2278 Right GGGTATAACTGCACATTCAGAGATT 2279 Left AAAAATCTCCAAAATATATGCCAAAG 2280 Right AACTGCACATTCAGAGATTCTTTCT EGFR Exon20-21 2 kb 2281 Left TTTCTACCAACTTCTGTCAAGCTCT 2282 Right TGCTATGTATTCTGTGGGTTAGACA 2283 Left TTTCTACCAACTTCTGTCAAGCTCT 2284 Right TGAGGTAATAAGTCAGCCATTTTTC 2285 Left TTTCTACCAACTTCTGTCAAGCTCT 2286 Right GGTAATAAGTCAGCCATTTTTCCTT 2287 Left AGAAAGAATCTCTGAATGTGCAGTT 2288 Right CAAAGTAACAATCAACAGACACTGG 2289 Left TTTCTACCAACTTCTGTCAAGCTCT 2290 Right ATAAAGGCCCATGTTCTCTTTACTT 2291 Left TTTCTACCAACTTCTGTCAAGCTCT 2292 Right CCTTCTTGGCTGTAAGATCAACTAA 2293 Left AATCTCTGAATGTGCAGTTATACCC 2294 Right CAAAGTAACAATCAACAGACACTGG 2295 Left TTTCTACCAACTTCTGTCAAGCTCT 2296 Right ATTACTCTCTGGCTTTTGTCCTTCT 2297 Left GTTTCTACCAACTTCTGTCAAGCTC 2298 Right TGAGGTAATAAGTCAGCCATTTTTC 2299 Left AGAAAGAATCTCTGAATGTGCAGTT 2300 Right AAAGATGAGATAACTTGGTGGAGTG 2301 Left AGAAAGAATCTCTGAATGTGCAGTT 2302 Right CAAAGATGAGATAACTTGGTGGAGT 2303 Left AGAAAGAATCTCTGAATGTGCAGTT 2304 Right TTTTCCAAGAACTTATTCCACAAAG 2305 Left AATCTCTGAATGTGCAGTTATACCC 2306 Right AAAGATGAGATAACTTGGTGGAGTG 2307 Left AATCTCTGAATGTGCAGTTATACCC 2308 Right CAAAGATGAGATAACTTGGTGGAGT 2309 Left TTTCTACCAACTTCTGTCAAGCTCT 2310 Right AATAAGTCAGCCATTTTTCCTTTTC 2311 Left TTTCTACCAACTTCTGTCAAGCTCT 2312 Right CTGCCCAGAGAAAATTAAACTGTAG 2313 Left AATCTCTGAATGTGCAGTTATACCC 2314 Right TTTTCCAAGAACTTATTCCACAAAG 2315 Left GTTTCTACCAACTTCTGTCAAGCTC 2316 Right GGTAATAAGTCAGCCATTTTTCCTT 2317 Left TGTGTTTCTACCAACTTCTGTCAAG 2318 Right TGCTATGTATTCTGTGGGTTAGACA 2319 Left GTTTCTACCAACTTCTGTCAAGCTC 2320 Right ATAAAGGCCCATGTTCTCTTTACTT EGFR Exon22 2 kb 2321 Left CACATAGCATTTGCACTGTATTAGG 2322 Right TTGAATCCAAAATAAAGGAATGTGT 2323 Left ATTTTGATATTTAAGGGAGGTCCTG 2324 Right TTCTCCACTACAAATCACCACAGTA 2325 Left CACATAGCATTTGCACTGTATTAGG 2326 Right TTCTCCACTACAAATCACCACAGTA 2327 Left GTCTGTAGGTTACACACAAATGCTG 2328 Right CACACTGAGCACTCAATAAAGAGAA 2329 Left CACATAGCATTTGCACTGTATTAGG 2330 Right ATTCTTCAAAGGTAGCTGATTGATG 2331 Left GTCTGTAGGTTACACACAAATGCTG 2332 Right TTCTCCACTACAAATCACCACAGTA 2333 Left AGGTAATCAGGAGATGCTGTAGATG 2334 Right CACACTGAGCACTCAATAAAGAGAA 2335 Left GTCTGTAGGTTACACACAAATGCTG 2336 Right ATTCTTCAAAGGTAGCTGATTGATG 2337 Left ATTTTGATATTTAAGGGAGGTCCTG 2338 Right GCTTACCTTGTTATCAAGTCCTGAA 2339 Left CACATAGCATTTGCACTGTATTAGG 2340 Right GCTTACCTTGTTATCAAGTCCTGAA 2341 Left AGGTAATCAGGAGATGCTGTAGATG 2342 Right ATTCTTCAAAGGTAGCTGATTGATG 2343 Left GTCTGTAGGTTACACACAAATGCTG 2344 Right GCTTACCTTGTTATCAAGTCCTGAA 2345 Left AGGTAATCAGGAGATGCTGTAGATG 2346 Right GCTTACCTTGTTATCAAGTCCTGAA 2347 Left GTATTTTGAAACTCAAGATCGCATT 2348 Right TTATACACATAGCGGAGTGATCAAA 2349 Left TCTGAGAAAGAGTCTGCTAAGGAAG 2350 Right TTGAATCCAAAATAAAGGAATGTGT 2351 Left CTCTGAGAAAGAGTCTGCTAAGGAA 2352 Right TTGAATCCAAAATAAAGGAATGTGT 2353 Left TCGGTACTGAACATATACGGACTTT 2354 Right TTGAATCCAAAATAAAGGAATGTGT 2355 Left TCGGTACTGAACATATACGGACTTT 2356 Right CACACTGAGCACTCAATAAAGAGAA 2357 Left CACATAGCATTTGCACTGTATTAGG 2358 Right CAAAGGTAGCTGATTGATGAGAGTT 2359 Left TCTGAGAAAGAGTCTGCTAAGGAAG 2360 Right ATCCAAAATAAAGGAATGTGTGTGT EGFR Exon18-21 5 kb 2361 Left ATGCCAAAGAAGTAGAATGAGAAAA 2362 Right TGCTATGTATTCTGTGGGTTAGACA 2363 Left ATGCCAAAGAAGTAGAATGAGAAAA 2364 Right CCTTCTTGGCTGTAAGATCAACTAA 2365 Left ATGCCAAAGAAGTAGAATGAGAAAA 2366 Right GTGCACTTAACTTTTAAGCCTTGAC 2367 Left ATGCCAAAGAAGTAGAATGAGAAAA 2368 Right AGATTGTAAGTGAAAGGCTTCACAG 2369 Left TCTCCAAAATATATGCCAAAGAAGT 2370 Right ATCTATCTTCTACCCCATTTCCAAC 2371 Left ATGCCAAAGAAGTAGAATGAGAAAA 2372 Right CTGCCCAGAGAAAATTAAACTGTAG 2373 Left TCTCCAAAATATATGCCAAAGAAGT 2374 Right TGCTATGTATTCTGTGGGTTAGACA 2375 Left TGCCAAAGAAGTAGAATGAGAAAAA 2376 Right ATCTATCTTCTACCCCATTTCCAAC 2377 Left TCTCCAAAATATATGCCAAAGAAGT 2378 Right TGAGGTAATAAGTCAGCCATTTTTC 2379 Left TGCCAAAGAAGTAGAATGAGAAAAA 2380 Right TGCTATGTATTCTGTGGGTTAGACA 2381 Left ATGCCAAAGAAGTAGAATGAGAAAA 2382 Right AGACATTTTTATAAAGGCCCATGTT 2383 Left ATGCCAAAGAAGTAGAATGAGAAAA 2384 Right ATTGTAAGTGAAAGGCTTCACAGAT 2385 Left TCTCCAAAATATATGCCAAAGAAGT 2386 Right GGTAATAAGTCAGCCATTTTTCCTT 2387 Left AATCTCCAAAATATATGCCAAAGAA 2388 Right ATCTATCTTCTACCCCATTTCCAAC 2389 Left AAATCTCCAAAATATATGCCAAAGA 2390 Right ATCTATCTTCTACCCCATTTCCAAC 2391 Left AAATCTCCAAAATATATGCCAAAGA 2392 Right TGCTATGTATTCTGTGGGTTAGACA 2393 Left TCTCCAAAATATATGCCAAAGAAGT 2394 Right CCTTCTTGGCTGTAAGATCAACTAA 2395 Left TCTCCAAAATATATGCCAAAGAAGT 2396 Right GTGCACTTAACTTTTAAGCCTTGAC 2397 Left AATCTCCAAAATATATGCCAAAGAA 2398 Right TGAGGTAATAAGTCAGCCATTTTTC 2399 Left AAATCTCCAAAATATATGCCAAAGA 2400 Right TGAGGTAATAAGTCAGCCATTTTTC EGFR Exon22 5 kb 2401 Left GTTGGAAATGGGGTAGAAGATAGAT 2402 Right TTGAATCCAAAATAAAGGAATGTGT 2403 Left GTTGGAAATGGGGTAGAAGATAGAT 2404 Right CACACTGAGCACTCAATAAAGAGAA 2405 Left ATTTTGATATTTAAGGGAGGTCCTG 2406 Right CTCTCCCATCAACATTTAGAAGAAA 2407 Left CACATAGCATTTGCACTGTATTAGG 2408 Right CTCTCCCATCAACATTTAGAAGAAA 2409 Left ATTTTGATATTTAAGGGAGGTCCTG 2410 Right TACAACAAACACAAGAATGGCTTTA 2411 Left CACATAGCATTTGCACTGTATTAGG 2412 Right TACAACAAACACAAGAATGGCTTTA 2413 Left GTTGGAAATGGGGTAGAAGATAGAT 2414 Right TTCTCCACTACAAATCACCACAGTA 2415 Left ATATCTGAATAAAAGGTCACCACCA 2416 Right CTCTCCCATCAACATTTAGAAGAAA 2417 Left CCATATCTGAATAAAAGGTCACCAC 2418 Right CTCTCCCATCAACATTTAGAAGAAA 2419 Left TATCTGAATAAAAGGTCACCACCAT 2420 Right CTCTCCCATCAACATTTAGAAGAAA 2421 Left CACATAGCATTTGCACTGTATTAGG 2422 Right GGGTCAAATAAACCTCCACTTATCT 2423 Left GTTGGAAATGGGGTAGAAGATAGAT 2424 Right ATTCTTCAAAGGTAGCTGATTGATG 2425 Left ATTTTGATATTTAAGGGAGGTCCTG 2426 Right TATAAGCCAATAAATCCCATTTTGA 2427 Left CACATAGCATTTGCACTGTATTAGG 2428 Right TATAAGCCAATAAATCCCATTTTGA 2429 Left GTCTGTAGGTTACACACAAATGCTG 2430 Right TACAACAAACACAAGAATGGCTTTA 2431 Left TACAGATTATGATGACTGCCTCAAA 2432 Right TACAACAAACACAAGAATGGCTTTA 2433 Left AGGAAAATAACACACACTCTCCTTG 2434 Right TTACTGGGAGATGATTAAGAACAGC 2435 Left GTCTGTAGGTTACACACAAATGCTG 2436 Right GGGTCAAATAAACCTCCACTTATCT 2437 Left ATATCTGAATAAAAGGTCACCACCA 2438 Right TATAAGCCAATAAATCCCATTTTGA 2439 Left CCATATCTGAATAAAAGGTCACCAC 2440 Right TATAAGCCAATAAATCCCATTTTGA

TABLE 9 KIT Capture Primer List for NGS Panel Seq. ID Primer Sequence KIT Exon8 150-175 bases 2441 Left ATATGGCCATTTCTGTTTTCCTGTA 2442 Right ATAAGCAGTGCCAAAAATAATCATC 2443 Left CTGACATATGGCCATTTCTGTTT 2444 Right ATAAGCAGTGCCAAAAATAATCATC 2445 Left GACATATGGCCATTTCTGTTTTC 2446 Right ATAAGCAGTGCCAAAAATAATCATC 2447 Left CTGACATATGGCCATTTCTGTTTT 2448 Right ATAAGCAGTGCCAAAAATAATCATC 2449 Left ATATGGCCATTTCTGTTTTCCTGTA 2450 Right GCATTATAAGCAGTGCCAAAAATAA 2451 Left TATGGCCATTTCTGTTTTCCTGTAG 2452 Right ATAAGCAGTGCCAAAAATAATCATC 2453 Left TGACATATGGCCATTTCTGTTTT 2454 Right ATAAGCAGTGCCAAAAATAATCATC 2455 Left ATATGGCCATTTCTGTTTTCCTGTA 2456 Right TTATAAGCAGTGCCAAAAATAATCA 2457 Left TATGGCCATTTCTGTTTTCCTGTA 2458 Right ATAAGCAGTGCCAAAAATAATCATC 2459 Left GACATATGGCCATTTCTGTTTTC 2460 Right TTATAAGCAGTGCCAAAAATAATCA 2461 Left CTGACATATGGCCATTTCTGTTTTC 2462 Right ATAAGCAGTGCCAAAAATAATCATC 2463 Left ATATGGCCATTTCTGTTTTCCTGTA 2464 Right TATAAGCAGTGCCAAAAATAATCATC 2465 Left GGCCATTTCTGTTTTCCTGTAG 2466 Right ATAAGCAGTGCCAAAAATAATCATC 2467 Left TATGGCCATTTCTGTTTTCCTGTAG 2468 Right GCATTATAAGCAGTGCCAAAAATAA 2469 Left ATATGGCCATTTCTGTTTTCCTGT 2470 Right ATAAGCAGTGCCAAAAATAATCATC 2471 Left ACATATGGCCATTTCTGTTTTCCT 2472 Right ATAAGCAGTGCCAAAAATAATCATC 2473 Left TATGGCCATTTCTGTTTTCCTGTAG 2474 Right TTATAAGCAGTGCCAAAAATAATCA 2475 Left TGACATATGGCCATTTCTGTTTTC 2476 Right ATAAGCAGTGCCAAAAATAATCATC 2477 Left ATATGGCCATTTCTGTTTTCCTGTA 2478 Right TAAGCAGTGCCAAAAATAATCATC 2479 Left GACATATGGCCATTTCTGTTTTC 2480 Right TATAAGCAGTGCCAAAAATAATCATC KIT Exon8 176-200 bases 2481 Left AGGTTTTCCAGCACTCTGACATA 2482 Right ATAAGCAGTGCCAAAAATAATCATC 2483 Left ACTCTGACATATGGCCATTTCTGTT 2484 Right ATAAGCAGTGCCAAAAATAATCATC 2485 Left CTGACATATGGCCATTTCTGTTT 2486 Right ATAAGCAGTGCCAAAAATAATCATC 2487 Left CTGACATATGGCCATTTCTGTTTT 2488 Right ATAAGCAGTGCCAAAAATAATCATC 2489 Left ATATGGCCATTTCTGTTTTCCTGTA 2490 Right GCATTATAAGCAGTGCCAAAAATAA 2491 Left CTCTGACATATGGCCATTTCTGTT 2492 Right ATAAGCAGTGCCAAAAATAATCATC 2493 Left CTCTGACATATGGCCATTTCTGTTT 2494 Right ATAAGCAGTGCCAAAAATAATCATC 2495 Left AGGTTTTCCAGCACTCTGACATA 2496 Right GCATTATAAGCAGTGCCAAAAATAA 2497 Left ACTCTGACATATGGCCATTTCTGTT 2498 Right GCATTATAAGCAGTGCCAAAAATAA 2499 Left TCTGACATATGGCCATTTCTGTT 2500 Right ATAAGCAGTGCCAAAAATAATCATC 2501 Left GACATATGGCCATTTCTGTTTTC 2502 Right GCATTATAAGCAGTGCCAAAAATAA 2503 Left AGGTTTTCCAGCACTCTGACATA 2504 Right TTATAAGCAGTGCCAAAAATAATCA 2505 Left ACTCTGACATATGGCCATTTCTGTT 2506 Right TTATAAGCAGTGCCAAAAATAATCA 2507 Left GAGGTTTTCCAGCACTCTGACATA 2508 Right ATAAGCAGTGCCAAAAATAATCATC 2509 Left CTGACATATGGCCATTTCTGTTT 2510 Right TTATAAGCAGTGCCAAAAATAATCA 2511 Left TCTGACATATGGCCATTTCTGTTT 2512 Right ATAAGCAGTGCCAAAAATAATCATC 2513 Left GACATATGGCCATTTCTGTTTTC 2514 Right TTATAAGCAGTGCCAAAAATAATCA 2515 Left CTGACATATGGCCATTTCTGTTTTC 2516 Right ATAAGCAGTGCCAAAAATAATCATC 2517 Left TCTGACATATGGCCATTTCTGTTTT 2518 Right ATAAGCAGTGCCAAAAATAATCATC 2519 Left CTGACATATGGCCATTTCTGTTTT 2520 Right TTATAAGCAGTGCCAAAAATAATCA KIT Exon8 201-300 bases 2521 Left TTAGAGAGGGAGTGAAGTGAATGTT 2522 Right ATAAGCAGTGCCAAAAATAATCATC 2523 Left GATTAGAGAGGGAGTGAAGTGAATG 2524 Right ATAAGCAGTGCCAAAAATAATCATC 2525 Left GGATTAGAGAGGGAGTGAAGTGAAT 2526 Right ATAAGCAGTGCCAAAAATAATCATC 2527 Left GTAGGGATTAGAGAGGGAGTGAAGT 2528 Right ATAAGCAGTGCCAAAAATAATCATC 2529 Left TTAGAGAGGGAGTGAAGTGAATGTT 2530 Right GCATTATAAGCAGTGCCAAAAATAA 2531 Left GATTAGAGAGGGAGTGAAGTGAATG 2532 Right GCATTATAAGCAGTGCCAAAAATAA 2533 Left CAGGAAGGTTGTAGGGATTAGAGAG 2534 Right ATAAGCAGTGCCAAAAATAATCATC 2535 Left CTCAGGAAGGTTGTAGGGATTAGAG 2536 Right ATAAGCAGTGCCAAAAATAATCATC 2537 Left GGATTAGAGAGGGAGTGAAGTGAAT 2538 Right GCATTATAAGCAGTGCCAAAAATAA 2539 Left GTAGGGATTAGAGAGGGAGTGAAGT 2540 Right GCATTATAAGCAGTGCCAAAAATAA 2541 Left GATTAGAGAGGGAGTGAAGTGAATG 2542 Right TTATAAGCAGTGCCAAAAATAATCA 2543 Left GGATTAGAGAGGGAGTGAAGTGAAT 2544 Right TTATAAGCAGTGCCAAAAATAATCA 2545 Left TCAGGAAGGTTGTAGGGATTAGAG 2546 Right ATAAGCAGTGCCAAAAATAATCATC 2547 Left CAGGAAGGTTGTAGGGATTAGAGA 2548 Right ATAAGCAGTGCCAAAAATAATCATC 2549 Left CTCAGGAAGGTTGTAGGGATTAGA 2550 Right ATAAGCAGTGCCAAAAATAATCATC 2551 Left TAGAGAGGGAGTGAAGTGAATGTTG 2552 Right ATAAGCAGTGCCAAAAATAATCATC 2553 Left GTAGGGATTAGAGAGGGAGTGAAGT 2554 Right TTATAAGCAGTGCCAAAAATAATCA 2555 Left AGGTTGTAGGGATTAGAGAGGGAGT 2556 Right ATAAGCAGTGCCAAAAATAATCATC 2557 Left ATTAGAGAGGGAGTGAAGTGAATGTT 2558 Right ATAAGCAGTGCCAAAAATAATCATC 2559 Left GGATTAGAGAGGGAGTGAAGTGAA 2560 Right ATAAGCAGTGCCAAAAATAATCATC KIT Exon8 301-400 bases 2561 Left TTAGAGAGGGAGTGAAGTGAATGTT 2562 Right TCATTCAGTAATGATTTTTCAGCAA 2563 Left TTAGAGAGGGAGTGAAGTGAATGTT 2564 Right TCAGTAATGATTTTTCAGCAAACAA 2565 Left GATTAGAGAGGGAGTGAAGTGAATG 2566 Right TCAGTAATGATTTTTCAGCAAACAA 2567 Left GATTAGAGAGGGAGTGAAGTGAATG 2568 Right TTCAGTAATGATTTTTCAGCAAACA 2569 Left GGATTAGAGAGGGAGTGAAGTGAAT 2570 Right TCAGTAATGATTTTTCAGCAAACAA 2571 Left GGATTAGAGAGGGAGTGAAGTGAAT 2572 Right TTCAGTAATGATTTTTCAGCAAACA 2573 Left TTAGAGAGGGAGTGAAGTGAATGTT 2574 Right AATTATCCCTTCTAAAAAGCCACAT 2575 Left GATTAGAGAGGGAGTGAAGTGAATG 2576 Right AATTATCCCTTCTAAAAAGCCACAT 2577 Left GGATTAGAGAGGGAGTGAAGTGAAT 2578 Right AATTATCCCTTCTAAAAAGCCACAT 2579 Left TTAGAGAGGGAGTGAAGTGAATGTT 2580 Right GTCATTCAGTAATGATTTTTCAGCA 2581 Left TTAGAGAGGGAGTGAAGTGAATGTT 2582 Right CAGCAAACAAAATTAATGTCTACCA 2583 Left GATTAGAGAGGGAGTGAAGTGAATG 2584 Right CAGCAAACAAAATTAATGTCTACCA 2585 Left GGATTAGAGAGGGAGTGAAGTGAAT 2586 Right CAGCAAACAAAATTAATGTCTACCA 2587 Left GTAGGGATTAGAGAGGGAGTGAAGT 2588 Right CAGCAAACAAAATTAATGTCTACCA 2589 Left TAGAGAGGGAGTGAAGTGAATGTTG 2590 Right TCATTCAGTAATGATTTTTCAGCAA 2591 Left ATTAGAGAGGGAGTGAAGTGAATGTT 2592 Right TCATTCAGTAATGATTTTTCAGCAA 2593 Left TAGAGAGGGAGTGAAGTGAATGTTG 2594 Right TCAGTAATGATTTTTCAGCAAACAA 2595 Left TAGAGAGGGAGTGAAGTGAATGTTG 2596 Right TTCAGTAATGATTTTTCAGCAAACA 2597 Left ATTAGAGAGGGAGTGAAGTGAATGTT 2598 Right TCAGTAATGATTTTTCAGCAAACAA 2599 Left CAGGAAGGTTGTAGGGATTAGAGAG 2600 Right AATTATCCCTTCTAAAAAGCCACAT KIT Exon8 401-500 bases 2601 Left GATTAGAGAGGGAGTGAAGTGAATG 2602 Right TCATTCAGTAATGATTTTTCAGCAA 2603 Left GGATTAGAGAGGGAGTGAAGTGAAT 2604 Right TCATTCAGTAATGATTTTTCAGCAA 2605 Left GTAGGGATTAGAGAGGGAGTGAAGT 2606 Right TCATTCAGTAATGATTTTTCAGCAA 2607 Left GTAGGGATTAGAGAGGGAGTGAAGT 2608 Right TCAGTAATGATTTTTCAGCAAACAA 2609 Left GTAGGGATTAGAGAGGGAGTGAAGT 2610 Right TTCAGTAATGATTTTTCAGCAAACA 2611 Left TTAGAGAGGGAGTGAAGTGAATGTT 2612 Right AAATTGCATGATAAATCCAGAAAGA 2613 Left TTAGAGAGGGAGTGAAGTGAATGTT 2614 Right GAAATTGCATGATAAATCCAGAAAG 2615 Left GATTAGAGAGGGAGTGAAGTGAATG 2616 Right AAATTGCATGATAAATCCAGAAAGA 2617 Left GATTAGAGAGGGAGTGAAGTGAATG 2618 Right GAAATTGCATGATAAATCCAGAAAG 2619 Left GGATTAGAGAGGGAGTGAAGTGAAT 2620 Right AAATTGCATGATAAATCCAGAAAGA 2621 Left GGATTAGAGAGGGAGTGAAGTGAAT 2622 Right GAAATTGCATGATAAATCCAGAAAG 2623 Left GTAGGGATTAGAGAGGGAGTGAAGT 2624 Right AAATTGCATGATAAATCCAGAAAGA 2625 Left GTAGGGATTAGAGAGGGAGTGAAGT 2626 Right GAAATTGCATGATAAATCCAGAAAG 2627 Left TTAGAAGCAGTCTTCAGATCCCTAC 2628 Right ATAAGCAGTGCCAAAAATAATCATC 2629 Left GATTAGAGAGGGAGTGAAGTGAATG 2630 Right GTCATTCAGTAATGATTTTTCAGCA 2631 Left GGATTAGAGAGGGAGTGAAGTGAAT 2632 Right GTCATTCAGTAATGATTTTTCAGCA 2633 Left AGATTTTTACCTGTGGAACACTTTG 2634 Right GCATTATAAGCAGTGCCAAAAATAA 2635 Left GTAGGGATTAGAGAGGGAGTGAAGT 2636 Right GTCATTCAGTAATGATTTTTCAGCA 2637 Left CAGGAAGGTTGTAGGGATTAGAGAG 2638 Right TCATTCAGTAATGATTTTTCAGCAA 2639 Left CTCAGGAAGGTTGTAGGGATTAGAG 2640 Right TCATTCAGTAATGATTTTTCAGCAA KIT Exon8 501-600 bases 2641 Left TTAGAGAGGGAGTGAAGTGAATGTT 2642 Right CGATCATTACTTTTTGGTAACTTGG 2643 Left GATTAGAGAGGGAGTGAAGTGAATG 2644 Right CGATCATTACTTTTTGGTAACTTGG 2645 Left GGATTAGAGAGGGAGTGAAGTGAAT 2646 Right CGATCATTACTTTTTGGTAACTTGG 2647 Left TTAGAGAGGGAGTGAAGTGAATGTT 2648 Right CATTACTTTTTGGTAACTTGGCAAT 2649 Left TTAGAGAGGGAGTGAAGTGAATGTT 2650 Right GCGATCATTACTTTTTGGTAACTTG 2651 Left TTAGAGAGGGAGTGAAGTGAATGTT 2652 Right TGCGATCATTACTTTTTGGTAACTT 2653 Left GATTAGAGAGGGAGTGAAGTGAATG 2654 Right CATTACTTTTTGGTAACTTGGCAAT 2655 Left GATTAGAGAGGGAGTGAAGTGAATG 2656 Right GCGATCATTACTTTTTGGTAACTTG 2657 Left GGATTAGAGAGGGAGTGAAGTGAAT 2658 Right CATTACTTTTTGGTAACTTGGCAAT 2659 Left ACTCCTAATTTCATCCATTCCAGTT 2660 Right ATAAGCAGTGCCAAAAATAATCATC 2661 Left AACTCCTAATTTCATCCATTCCAGT 2662 Right ATAAGCAGTGCCAAAAATAATCATC 2663 Left GTAGGGATTAGAGAGGGAGTGAAGT 2664 Right CATTACTTTTTGGTAACTTGGCAAT 2665 Left GGCAGGAATCCTTTAAAGTAGATTT 2666 Right ATAAGCAGTGCCAAAAATAATCATC 2667 Left TTAGAAGCAGTCTTCAGATCCCTAC 2668 Right AATTATCCCTTCTAAAAAGCCACAT 2669 Left TTAGAGAGGGAGTGAAGTGAATGTT 2670 Right GATAACTACAGTCACATTTCCCACA 2671 Left AACAACTCCTAATTTCATCCATTCC 2672 Right ATAAGCAGTGCCAAAAATAATCATC 2673 Left GATTAGAGAGGGAGTGAAGTGAATG 2674 Right GATAACTACAGTCACATTTCCCACA 2675 Left TTAGAGAGGGAGTGAAGTGAATGTT 2676 Right TAACTACAGTCACATTTCCCACACA 2677 Left GGATTAGAGAGGGAGTGAAGTGAAT 2678 Right GATAACTACAGTCACATTTCCCACA 2679 Left CACTTTGGAGTCCTAGAGTTTGATT 2680 Right AATTATCCCTTCTAAAAAGCCACAT KIT Exon8 601-800 bases 2681 Left AGATTTTTACCTGTGGAACACTTTG 2682 Right TCATTCAGTAATGATTTTTCAGCAA 2683 Left AGATTTTTACCTGTGGAACACTTTG 2684 Right TTCAGTAATGATTTTTCAGCAAACA 2685 Left AGATTTTTACCTGTGGAACACTTTG 2686 Right TCAGTAATGATTTTTCAGCAAACAA 2687 Left GGAGAAAATTCATGTAAGAGCAAAA 2688 Right ATAAGCAGTGCCAAAAATAATCATC 2689 Left AATTCATGTAAGAGCAAAAGAGTGG 2690 Right ATAAGCAGTGCCAAAAATAATCATC 2691 Left GGAGAAAATTCATGTAAGAGCAAAA 2692 Right AATTATCCCTTCTAAAAAGCCACAT 2693 Left AGATTTTTACCTGTGGAACACTTTG 2694 Right AAATTGCATGATAAATCCAGAAAGA 2695 Left AGATTTTTACCTGTGGAACACTTTG 2696 Right GAAATTGCATGATAAATCCAGAAAG 2697 Left AATTCATGTAAGAGCAAAAGAGTGG 2698 Right AATTATCCCTTCTAAAAAGCCACAT 2699 Left GGGCTTATCTTTTCCTCTAACAACT 2700 Right TCATTCAGTAATGATTTTTCAGCAA 2701 Left GGGCTTATCTTTTCCTCTAACAACT 2702 Right TCAGTAATGATTTTTCAGCAAACAA 2703 Left GGGCTTATCTTTTCCTCTAACAACT 2704 Right TTCAGTAATGATTTTTCAGCAAACA 2705 Left AGATTTTTACCTGTGGAACACTTTG 2706 Right AATTATCCCTTCTAAAAAGCCACAT 2707 Left AGGCTGGTTTTCTTTTCTAGTTTTC 2708 Right ATAAGCAGTGCCAAAAATAATCATC 2709 Left GGCTGGTTTTCTTTTCTAGTTTTCT 2710 Right ATAAGCAGTGCCAAAAATAATCATC 2711 Left ACTCCTAATTTCATCCATTCCAGTT 2712 Right TCATTCAGTAATGATTTTTCAGCAA 2713 Left AACTCCTAATTTCATCCATTCCAGT 2714 Right TCATTCAGTAATGATTTTTCAGCAA 2715 Left AGGAGAAAATTCATGTAAGAGCAAA 2716 Right ATAAGCAGTGCCAAAAATAATCATC 2717 Left AAGGAGAAAATTCATGTAAGAGCAA 2718 Right ATAAGCAGTGCCAAAAATAATCATC 2719 Left AAAGGAGAAAATTCATGTAAGAGCA 2720 Right ATAAGCAGTGCCAAAAATAATCATC KIT Exon8 801-1000 bases 2721 Left GGAGAAAATTCATGTAAGAGCAAAA 2722 Right TCATTCAGTAATGATTTTTCAGCAA 2723 Left GGAGAAAATTCATGTAAGAGCAAAA 2724 Right TCAGTAATGATTTTTCAGCAAACAA 2725 Left GGAGAAAATTCATGTAAGAGCAAAA 2726 Right TTCAGTAATGATTTTTCAGCAAACA 2727 Left AATTCATGTAAGAGCAAAAGAGTGG 2728 Right TCATTCAGTAATGATTTTTCAGCAA 2729 Left AATTCATGTAAGAGCAAAAGAGTGG 2730 Right TCAGTAATGATTTTTCAGCAAACAA 2731 Left AATTCATGTAAGAGCAAAAGAGTGG 2732 Right TTCAGTAATGATTTTTCAGCAAACA 2733 Left TTAGAGCATTTCTGCTGTTACAGTG 2734 Right ATAAGCAGTGCCAAAAATAATCATC 2735 Left GGAGAAAATTCATGTAAGAGCAAAA 2736 Right AAATTGCATGATAAATCCAGAAAGA 2737 Left GGAGAAAATTCATGTAAGAGCAAAA 2738 Right GAAATTGCATGATAAATCCAGAAAG 2739 Left ATACCAAATTAGAGCATTTCTGCTG 2740 Right ATAAGCAGTGCCAAAAATAATCATC 2741 Left AGGCTGGTTTTCTTTTCTAGTTTTC 2742 Right TCATTCAGTAATGATTTTTCAGCAA 2743 Left GGCTGGTTTTCTTTTCTAGTTTTCT 2744 Right TCATTCAGTAATGATTTTTCAGCAA 2745 Left AATTCATGTAAGAGCAAAAGAGTGG 2746 Right AAATTGCATGATAAATCCAGAAAGA 2747 Left AATTCATGTAAGAGCAAAAGAGTGG 2748 Right GAAATTGCATGATAAATCCAGAAAG 2749 Left AGATTTTTACCTGTGGAACACTTTG 2750 Right TGTTGTAATTGTGCGATCATTACTT 2751 Left AGGCTGGTTTTCTTTTCTAGTTTTC 2752 Right TTCAGTAATGATTTTTCAGCAAACA 2753 Left AGATTTTTACCTGTGGAACACTTTG 2754 Right CGATCATTACTTTTTGGTAACTTGG 2755 Left TTAGAGAGGGAGTGAAGTGAATGTT 2756 Right GAACCCTACTTAGTATTCCCCAAAA 2757 Left AGGAGAAAATTCATGTAAGAGCAAA 2758 Right TCATTCAGTAATGATTTTTCAGCAA 2759 Left AAGGAGAAAATTCATGTAAGAGCAA 2760 Right TCATTCAGTAATGATTTTTCAGCAA KIT Exon8 2 kb 2761 Left GGAGAAAATTCATGTAAGAGCAAAA 2762 Right GGACTGAAGTTTGAGTTCTAAGCAG 2763 Left GGAGAAAATTCATGTAAGAGCAAAA 2764 Right AGGACTGAAGTTTGAGTTCTAAGCA 2765 Left TTAGAGCATTTCTGCTGTTACAGTG 2766 Right CCTGTTTCCTTTCTTAACACCTACA 2767 Left GGAGAAAATTCATGTAAGAGCAAAA 2768 Right CCTGTTTCCTTTCTTAACACCTACA 2769 Left AATTCATGTAAGAGCAAAAGAGTGG 2770 Right ACTGAAGTTTGAGTTCTAAGCAGGA 2771 Left AATTCATGTAAGAGCAAAAGAGTGG 2772 Right GGACTGAAGTTTGAGTTCTAAGCAG 2773 Left AATTCATGTAAGAGCAAAAGAGTGG 2774 Right AGGACTGAAGTTTGAGTTCTAAGCA 2775 Left GGAGAAAATTCATGTAAGAGCAAAA 2776 Right GACCAGAAAATAGTCAAAGTGAGGA 2777 Left TTCTGATCTGTCAGTCTTTCCTTCT 2778 Right TCATTCAGTAATGATTTTTCAGCAA 2779 Left ATACCAAATTAGAGCATTTCTGCTG 2780 Right CCTGTTTCCTTTCTTAACACCTACA 2781 Left TTAGAGCATTTCTGCTGTTACAGTG 2782 Right GCTATTCCCTGTTTCCTTTCTTAAC 2783 Left ATAAGTGCATCTTCCTTTCACTTTG 2784 Right GAACCCTACTTAGTATTCCCCAAAA 2785 Left GGAGAAAATTCATGTAAGAGCAAAA 2786 Right GCTATTCCCTGTTTCCTTTCTTAAC 2787 Left TTCTGATCTGTCAGTCTTTCCTTCT 2788 Right TTCAGTAATGATTTTTCAGCAAACA 2789 Left TTCTGATCTGTCAGTCTTTCCTTCT 2790 Right TCAGTAATGATTTTTCAGCAAACAA 2791 Left AATTCATGTAAGAGCAAAAGAGTGG 2792 Right GACCAGAAAATAGTCAAAGTGAGGA 2793 Left TTCCTTCTCACTGCATATATTTTCC 2794 Right TCATTCAGTAATGATTTTTCAGCAA 2795 Left ATACCAAATTAGAGCATTTCTGCTG 2796 Right GCTATTCCCTGTTTCCTTTCTTAAC 2797 Left TGAAATTGGTTATCCAAGAAAGGTA 2798 Right TCATTCAGTAATGATTTTTCAGCAA 2799 Left AGATTTTTACCTGTGGAACACTTTG 2800 Right ACTGAAGTTTGAGTTCTAAGCAGGA KIT Exon8-9 5 kb 2801 Left AGTTGCCCATGATAATTAAATGAAA 2802 Right AGGCAGTGTTAACTTTTGGATACAG 2803 Left CCCATGATAATTAAATGAAACTTGC 2804 Right AGGCAGTGTTAACTTTTGGATACAG 2805 Left GCCCATGATAATTAAATGAAACTTG 2806 Right AGGCAGTGTTAACTTTTGGATACAG 2807 Left TGCCCATGATAATTAAATGAAACTT 2808 Right AGGCAGTGTTAACTTTTGGATACAG 2809 Left TTGCCCATGATAATTAAATGAAACT 2810 Right AGGCAGTGTTAACTTTTGGATACAG 2811 Left AGTTTAGGCTTGCTTAGAAAAGGAG 2812 Right AGGCAGTGTTAACTTTTGGATACAG 2813 Left AGAGTTTAGGCTTGCTTAGAAAAGG 2814 Right AGGCAGTGTTAACTTTTGGATACAG 2815 Left GATTTCTTGTGTCGTGTCCTACTTT 2816 Right AGGCAGTGTTAACTTTTGGATACAG 2817 Left GCCCATGATAATTAAATGAAACTTG 2818 Right GCTTCCTTTATGGACGGTTTATATT 2819 Left TTGCCCATGATAATTAAATGAAACT 2820 Right GCTTCCTTTATGGACGGTTTATATT 2821 Left AGTTGCCCATGATAATTAAATGAAA 2822 Right GCTTCCTTTATGGACGGTTTATATT 2823 Left CCCATGATAATTAAATGAAACTTGC 2824 Right GCTTCCTTTATGGACGGTTTATATT 2825 Left TGCCCATGATAATTAAATGAAACTT 2826 Right GCTTCCTTTATGGACGGTTTATATT 2827 Left TTGCCCATGATAATTAAATGAAACT 2828 Right CCCCTTAAATTGGATTAAAAAGAAA 2829 Left AGTTGCCCATGATAATTAAATGAAA 2830 Right CCCCTTAAATTGGATTAAAAAGAAA 2831 Left GCCCATGATAATTAAATGAAACTTG 2832 Right CCCCTTAAATTGGATTAAAAAGAAA 2833 Left CCCATGATAATTAAATGAAACTTGC 2834 Right CCCCTTAAATTGGATTAAAAAGAAA 2835 Left TGCCCATGATAATTAAATGAAACTT 2836 Right CCCCTTAAATTGGATTAAAAAGAAA 2837 Left AGTTTAGGCTTGCTTAGAAAAGGAG 2838 Right CCCCTTAAATTGGATTAAAAAGAAA 2839 Left AGAGTTTAGGCTTGCTTAGAAAAGG 2840 Right CCCCTTAAATTGGATTAAAAAGAAA KIT Exon9 200-250 bases 2841 Left GGCTTTTGTTTTCTTCCCTTTAG 2842 Right CATCCCCTTAAATTGGATTAAAAAG 2843 Left GGCTTTTGTTTTCTTCCCTTTAG 2844 Right ATCCCCTTAAATTGGATTAAAAAG 2845 Left GGGCTTTTGTTTTCTTCCCTTTAG 2846 Right ATCCCCTTAAATTGGATTAAAAAG 2847 Left GGCTTTTGTTTTCTTCCCTTTAG 2848 Right TCCCCTTAAATTGGATTAAAAAG 2849 Left GCTTTTGTTTTCTTCCCTTTAG 2850 Right CATCCCCTTAAATTGGATTAAAAAG 2851 Left AGGGCTTTTGTTTTCTTCCCTTTAG 2852 Right TCCCCTTAAATTGGATTAAAAAG 2853 Left GGGCTTTTGTTTTCTTCCCTTTAG 2854 Right TCCCCTTAAATTGGATTAAAAAG 2855 Left GCTTTTGTTTTCTTCCCTTTAG 2856 Right ACATCCCCTTAAATTGGATTAAAAAG 2857 Left GGCTTTTGTTTTCTTCCCTTTAG 2858 Right CCCCTTAAATTGGATTAAAAAG 2859 Left GCTTTTGTTTTCTTCCCTTTAG 2860 Right ATCCCCTTAAATTGGATTAAAAAG 2861 Left AGGGCTTTTGTTTTCTTCCCTTTAG 2862 Right CCCCTTAAATTGGATTAAAAAG 2863 Left GGGCTTTTGTTTTCTTCCCTTTAG 2864 Right CCCCTTAAATTGGATTAAAAAG 2865 Left GCTTTTGTTTTCTTCCCTTTAG 2866 Right TCCCCTTAAATTGGATTAAAAAG 2867 Left CTTTTGTTTTCTTCCCTTTAG 2868 Right CATCCCCTTAAATTGGATTAAAAAG 2869 Left CAGGGCTTTTGTTTTCTTCC 2870 Right CCCCTTAAATTGGATTAAAAAGAAAT ATAC 2871 Left CTTTTGTTTTCTTCCCTTTAG 2872 Right ACATCCCCTTAAATTGGATTAAAAAG 2873 Left CAGGGCTTTTGTTTTCTTCCCTTTAG 2874 Right CCCCTTAAATTGGATTAAAAAG 2875 Left CAGGGCTTTTGTTTTCTTCC 2876 Right CCCCTTAAATTGGATTAAAAAG 2877 Left GGCTTTTGTTTTCTTCCCTTTAG 2878 Right CATCCCCTTAAATTGGATTA 2879 Left GCTTTTGTTTTCTTCCCTTTAG 2880 Right CCCCTTAAATTGGATTAAAAAG KIT Exon9 251-300 bases 2881 Left CCACATCCCAAGTGTTTTATGTATT 2882 Right CCCCTTAAATTGGATTAAAAAGAAA 2883 Left CCACATCCCAAGTGTTTTATGTATT 2884 Right CATCCCCTTAAATTGGATTAAAAAG 2885 Left CCACATCCCAAGTGTTTTATGTATT 2886 Right TCCCCTTAAATTGGATTAAAAAGAA 2887 Left CCACATCCCAAGTGTTTTATGTATT 2888 Right ATCCCCTTAAATTGGATTAAAAAGA 2889 Left CCACATCCCAAGTGTTTTATGTATT 2890 Right CCCCTTAAATTGGATTAAAAAGAAAT 2891 Left CCACATCCCAAGTGTTTTATGTATT 2892 Right ATCCCCTTAAATTGGATTAAAAAGAA 2893 Left CCACATCCCAAGTGTTTTATGTAT 2894 Right CCCCTTAAATTGGATTAAAAAGAAA 2895 Left CCACATCCCAAGTGTTTTATGTATT 2896 Right TCCCCTTAAATTGGATTAAAAAGA 2897 Left CCACATCCCAAGTGTTTTATGTAT 2898 Right CATCCCCTTAAATTGGATTAAAAAG 2899 Left CCACATCCCAAGTGTTTTATGTAT 2900 Right TCCCCTTAAATTGGATTAAAAAGAA 2901 Left GCCACATCCCAAGTGTTTTATGTAT 2902 Right CCCCTTAAATTGGATTAAAAAGAAA 2903 Left CCACATCCCAAGTGTTTTATGTAT 2904 Right ATCCCCTTAAATTGGATTAAAAAGA 2905 Left CCACATCCCAAGTGTTTTATGTATT 2906 Right CCCCTTAAATTGGATTAAAAAGAA 2907 Left GGCTTTTGTTTTCTTCCCTTTAG 2908 Right TATGGTAGACAGAGCCTAAACATCC 2909 Left CTAGAGTAAGCCAGGGCTTTTGTTT 2910 Right TATGGTAGACAGAGCCTAAACATCC 2911 Left CTAGAGTAAGCCAGGGCTTTTGTT 2912 Right TATGGTAGACAGAGCCTAAACATCC 2913 Left CTAGAGTAAGCCAGGGCTTTTGTTT 2914 Right AACATCCCCTTAAATTGGATTAAAA 2915 Left CTAGAGTAAGCCAGGGCTTTTGTT 2916 Right AACATCCCCTTAAATTGGATTAAAA 2917 Left CCACATCCCAAGTGTTTTATGTATT 2918 Right TCCCCTTAAATTGGATTAAAAAGAAA 2919 Left GGCTTTTGTTTTCTTCCCTTTAG 2920 Right TCATGACTGATATGGTAGACAGAGC KIT Exon9 301-400 bases 2921 Left CTCACTAGGTCACCAAAGTGCTTAT 2922 Right CCCCTTAAATTGGATTAAAAAGAAA 2923 Left CTCACTAGGTCACCAAAGTGCTTAT 2924 Right AACATCCCCTTAAATTGGATTAAAA 2925 Left CCACATCCCAAGTGTTTTATGTATT 2926 Right TATGGTAGACAGAGCCTAAACATCC 2927 Left CTCACTAGGTCACCAAAGTGCTTAT 2928 Right CATCCCCTTAAATTGGATTAAAAAG 2929 Left CCACATCCCAAGTGTTTTATGTATT 2930 Right GTGATGCATGTATTACCAGAAATGA 2931 Left CCACATCCCAAGTGTTTTATGTATT 2932 Right AACATCCCCTTAAATTGGATTAAAA 2933 Left CACTAGGTCACCAAAGTGCTTATTC 2934 Right CCCCTTAAATTGGATTAAAAAGAAA 2935 Left TCACTAGGTCACCAAAGTGCTTATT 2936 Right CCCCTTAAATTGGATTAAAAAGAAA 2937 Left CTCACTAGGTCACCAAAGTGCTTAT 2938 Right TCCCCTTAAATTGGATTAAAAAGAA 2939 Left TTTGTTTTAAAAGTATGCCACATCC 2940 Right CCCCTTAAATTGGATTAAAAAGAAA 2941 Left TTTGTTTTAAAAGTATGCCACATCC 2942 Right TATGGTAGACAGAGCCTAAACATCC 2943 Left CTCACTAGGTCACCAAAGTGCTTAT 2944 Right ATCCCCTTAAATTGGATTAAAAAGA 2945 Left CCACATCCCAAGTGTTTTATGTATT 2946 Right TCATGACTGATATGGTAGACAGAGC 2947 Left CACTAGGTCACCAAAGTGCTTATTC 2948 Right AACATCCCCTTAAATTGGATTAAAA 2949 Left TCACTAGGTCACCAAAGTGCTTATT 2950 Right AACATCCCCTTAAATTGGATTAAAA 2951 Left TTTGTTTTAAAAGTATGCCACATCC 2952 Right GTGATGCATGTATTACCAGAAATGA 2953 Left TCACCAAAGTGCTTATTCTTAGACA 2954 Right CCCCTTAAATTGGATTAAAAAGAAA 2955 Left ACTCACTAGGTCACCAAAGTGCTTA 2956 Right CCCCTTAAATTGGATTAAAAAGAAA 2957 Left TTTGTTTTAAAAGTATGCCACATCC 2958 Right AACATCCCCTTAAATTGGATTAAAA 2959 Left AGGTCACCAAAGTGCTTATTCTTAG 2960 Right CCCCTTAAATTGGATTAAAAAGAAA KIT Exon9 401-500 bases 2961 Left CTCACTAGGTCACCAAAGTGCTTAT 2962 Right TATGGTAGACAGAGCCTAAACATCC 2963 Left TACAGTCGTAGAAACTCAGTGTTGG 2964 Right CCCCTTAAATTGGATTAAAAAGAAA 2965 Left CTCACTAGGTCACCAAAGTGCTTAT 2966 Right GTGATGCATGTATTACCAGAAATGA 2967 Left CTCACTAGGTCACCAAAGTGCTTAT 2968 Right TCATGACTGATATGGTAGACAGAGC 2969 Left TACAGTCGTAGAAACTCAGTGTTGG 2970 Right CATCCCCTTAAATTGGATTAAAAAG 2971 Left CACTAGGTCACCAAAGTGCTTATTC 2972 Right TATGGTAGACAGAGCCTAAACATCC 2973 Left TCACTAGGTCACCAAAGTGCTTATT 2974 Right TATGGTAGACAGAGCCTAAACATCC 2975 Left ACAGTCGTAGAAACTCAGTGTTGGT 2976 Right CCCCTTAAATTGGATTAAAAAGAAA 2977 Left TACAGTCGTAGAAACTCAGTGTTGG 2978 Right TCCCCTTAAATTGGATTAAAAAGAA 2979 Left CTCACTAGGTCACCAAAGTGCTTAT 2980 Right GGTCAATGTTGGAATGAACTTAAAA 2981 Left CACTAGGTCACCAAAGTGCTTATTC 2982 Right GTGATGCATGTATTACCAGAAATGA 2983 Left TCACTAGGTCACCAAAGTGCTTATT 2984 Right GTGATGCATGTATTACCAGAAATGA 2985 Left TACAGTCGTAGAAACTCAGTGTTGG 2986 Right ATCCCCTTAAATTGGATTAAAAAGA 2987 Left TCACCAAAGTGCTTATTCTTAGACA 2988 Right TATGGTAGACAGAGCCTAAACATCC 2989 Left ACTCACTAGGTCACCAAAGTGCTTA 2990 Right TATGGTAGACAGAGCCTAAACATCC 2991 Left ACAGTCGTAGAAACTCAGTGTTGGT 2992 Right AACATCCCCTTAAATTGGATTAAAA 2993 Left CTAGGTCACCAAAGTGCTTATTCTT 2994 Right TATGGTAGACAGAGCCTAAACATCC 2995 Left AGGTCACCAAAGTGCTTATTCTTAG 2996 Right TATGGTAGACAGAGCCTAAACATCC 2997 Left TCACCAAAGTGCTTATTCTTAGACA 2998 Right GTGATGCATGTATTACCAGAAATGA 2999 Left CACTAGGTCACCAAAGTGCTTATTC 3000 Right TCATGACTGATATGGTAGACAGAGC KIT Exon9 501-600 bases 3001 Left CCTCTATGCTATTTCTTTTCAACCA 3002 Right TATGGTAGACAGAGCCTAAACATCC 3003 Left CTCACTAGGTCACCAAAGTGCTTAT 3004 Right AGGCAGTGTTAACTTTTGGATACAG 3005 Left TTTTATGCTTTCCTCCTCTATGCTA 3006 Right CCCCTTAAATTGGATTAAAAAGAAA 3007 Left CTCACTAGGTCACCAAAGTGCTTAT 3008 Right GGCAGTGTTAACTTTTGGATACAGT 3009 Left GCTTTCCTCCTCTATGCTATTTCTT 3010 Right TATGGTAGACAGAGCCTAAACATCC 3011 Left TTTTATGCTTTCCTCCTCTATGCTA 3012 Right TATGGTAGACAGAGCCTAAACATCC 3013 Left CCACATCCCAAGTGTTTTATGTATT 3014 Right AGGCAGTGTTAACTTTTGGATACAG 3015 Left TCCTCCTCTATGCTATTTCTTTTCA 3016 Right TATGGTAGACAGAGCCTAAACATCC 3017 Left ATTTTATTGAATTCCTTTCCAATCC 3018 Right GTGATGCATGTATTACCAGAAATGA 3019 Left CTCACTAGGTCACCAAAGTGCTTAT 3020 Right GTAAATATATTCCCCCATTTGCTTT 3021 Left GCTTTCCTCCTCTATGCTATTTCTT 3022 Right AACATCCCCTTAAATTGGATTAAAA 3023 Left TTTTATGCTTTCCTCCTCTATGCTA 3024 Right AACATCCCCTTAAATTGGATTAAAA 3025 Left CCACATCCCAAGTGTTTTATGTATT 3026 Right GGCAGTGTTAACTTTTGGATACAGT 3027 Left TTTAGTAGAGACGAGGTTTCACCAT 3028 Right CCCCTTAAATTGGATTAAAAAGAAA 3029 Left GCTGAGATTACAGGTGTGAGCTACT 3030 Right CCCCTTAAATTGGATTAAAAAGAAA 3031 Left CTCACTAGGTCACCAAAGTGCTTAT 3032 Right GATTGTTCTAATTCTGTTTGGGTGT 3033 Left TACAGTCGTAGAAACTCAGTGTTGG 3034 Right GTAAATATATTCCCCCATTTGCTTT 3035 Left GCTGAGATTACAGGTGTGAGCTACT 3036 Right TATGGTAGACAGAGCCTAAACATCC 3037 Left CACTAGGTCACCAAAGTGCTTATTC 3038 Right AGGCAGTGTTAACTTTTGGATACAG 3039 Left TCACTAGGTCACCAAAGTGCTTATT 3040 Right AGGCAGTGTTAACTTTTGGATACAG KIT Exon9 801-1000 bases 3041 Left ATTCCTTTCCAATCCTTTCAGTAAC 3042 Right AGGCAGTGTTAACTTTTGGATACAG 3043 Left ATTTTATTGAATTCCTTTCCAATCC 3044 Right AGGCAGTGTTAACTTTTGGATACAG 3045 Left TACAGTCGTAGAAACTCAGTGTTGG 3046 Right AGGCAGTGTTAACTTTTGGATACAG 3047 Left TACATCCTTGATTTTGTTGTTGTTG 3048 Right CCCCTTAAATTGGATTAAAAAGAAA 3049 Left ATTCCTTTCCAATCCTTTCAGTAAC 3050 Right GGCAGTGTTAACTTTTGGATACAGT 3051 Left CCTCTATGCTATTTCTTTTCAACCA 3052 Right GTAAATATATTCCCCCATTTGCTTT 3053 Left ATTTTATTGAATTCCTTTCCAATCC 3054 Right GGCAGTGTTAACTTTTGGATACAGT 3055 Left TTCTGGTCTACATCCTTGATTTTGT 3056 Right CCCCTTAAATTGGATTAAAAAGAAA 3057 Left TACAGTCGTAGAAACTCAGTGTTGG 3058 Right GGCAGTGTTAACTTTTGGATACAGT 3059 Left CCTCTATGCTATTTCTTTTCAACCA 3060 Right GATTGTTCTAATTCTGTTTGGGTGT 3061 Left CCCTGTTTTACAGTCGTAGAAACTC 3062 Right AGGCAGTGTTAACTTTTGGATACAG 3063 Left ATTCCTTTCCAATCCTTTCAGTAAC 3064 Right GTAAATATATTCCCCCATTTGCTTT 3065 Left TACATCCTTGATTTTGTTGTTGTTG 3066 Right AACATCCCCTTAAATTGGATTAAAA 3067 Left AACCCTCTGCAATGGGTATTACTAT 3068 Right AGGCAGTGTTAACTTTTGGATACAG 3069 Left TTATTGAATTCCTTTCCAATCCTTT 3070 Right AGGCAGTGTTAACTTTTGGATACAG 3071 Left TTTTATTGAATTCCTTTCCAATCCT 3072 Right AGGCAGTGTTAACTTTTGGATACAG 3073 Left TTTATTGAATTCCTTTCCAATCCTT 3074 Right AGGCAGTGTTAACTTTTGGATACAG 3075 Left ATTTTATTGAATTCCTTTCCAATCC 3076 Right GTAAATATATTCCCCCATTTGCTTT 3077 Left CCTCTATGCTATTTCTTTTCAACCA 3078 Right TGCTTTCTCTAGCTCTTTTTAATGG 3079 Left CCACATCCCAAGTGTTTTATGTATT 3080 Right ACTACTCAAAACCTGAGAAAACACG KIT Exon9 2 kb 3081 Left TGTAGGTGTTAAGAAAGGAAACAGG 3082 Right GCTTCCTTTATGGACGGTTTATATT 3083 Left TCCTCACTTTGACTATTTTCTGGTC 3084 Right GCTTCCTTTATGGACGGTTTATATT 3085 Left GTTAAGAAAGGAAACAGGGAATAGC 3086 Right GCTTCCTTTATGGACGGTTTATATT 3087 Left TGTAGGTGTTAAGAAAGGAAACAGG 3088 Right ACTACTCAAAACCTGAGAAAACACG 3089 Left AAGTAATGATCGCACAATTACAACA 3090 Right CCCCTTAAATTGGATTAAAAAGAAA 3091 Left CCAAGTTACCAAAAAGTAATGATCG 3092 Right CCCCTTAAATTGGATTAAAAAGAAA 3093 Left TGGATAAGCTTGTTCTAGTGGGTAG 3094 Right ACTACTCAAAACCTGAGAAAACACG 3095 Left TGTAGGTGTTAAGAAAGGAAACAGG 3096 Right CCTCACTACTCAAAACCTGAGAAAA 3097 Left ATAACTAGGCCTTCCTGCTTAGAAC 3098 Right GCTTCCTTTATGGACGGTTTATATT 3099 Left GTTAAGAAAGGAAACAGGGAATAGC 3100 Right CCTCACTACTCAAAACCTGAGAAAA 3101 Left TGGATAAGCTTGTTCTAGTGGGTAG 3102 Right CCTCACTACTCAAAACCTGAGAAAA 3103 Left TCTGGTCTACATCCTTGATTTTGTT 3104 Right GCTTCCTTTATGGACGGTTTATATT 3105 Left TTCTGGTCTACATCCTTGATTTTGT 3106 Right GCTTCCTTTATGGACGGTTTATATT 3107 Left GGTCTACATCCTTGATTTTGTTGTT 3108 Right GCTTCCTTTATGGACGGTTTATATT 3109 Left GGAATAAGCCTCTTTATCACAACAA 3110 Right GTAAATATATTCCCCCATTTGCTTT 3111 Left TGTAGGTGTTAAGAAAGGAAACAGG 3112 Right TCTTTAAGCTTTCCTGTATTTTCCA 3113 Left AAGTAATGATCGCACAATTACAACA 3114 Right AACATCCCCTTAAATTGGATTAAAA 3115 Left TAACTAGGCCTTCCTGCTTAGAACT 3116 Right GCTTCCTTTATGGACGGTTTATATT 3117 Left TTCATGGAATAAGCCTCTTTATCAC 3118 Right GTAAATATATTCCCCCATTTGCTTT 3119 Left TTCTTTTGGGGAATACTAAGTAGGG 3120 Right GTAAATATATTCCCCCATTTGCTTT KIT Exon9-10 2 kb 3121 Left CCTCTATGCTATTTCTTTTCAACCA 3122 Right ATTAGAGCACTCTGGAGAGAGAACA 3123 Left ATTCCTTTCCAATCCTTTCAGTAAC 3124 Right AGCACTCTGGAGAGAGAACAAATAA 3125 Left ATTCCTTTCCAATCCTTTCAGTAAC 3126 Right ATTAGAGCACTCTGGAGAGAGAACA 3127 Left ATTTTATTGAATTCCTTTCCAATCC 3128 Right ATTAGAGCACTCTGGAGAGAGAACA 3129 Left GCTTTCCTCCTCTATGCTATTTCTT 3130 Right ATTAGAGCACTCTGGAGAGAGAACA 3131 Left AACCCTCTGCAATGGGTATTACTAT 3132 Right AGCACTCTGGAGAGAGAACAAATAA 3133 Left GCTGAGATTACAGGTGTGAGCTACT 3134 Right AGCACTCTGGAGAGAGAACAAATAA 3135 Left ATTCCTTTCCAATCCTTTCAGTAAC 3136 Right GCACTCTGGAGAGAGAACAAATAAA 3137 Left ATTCCTTTCCAATCCTTTCAGTAAC 3138 Right CTCTGGAGAGAGAACAAATAAATGG 3139 Left AACCCTCTGCAATGGGTATTACTAT 3140 Right ATTAGAGCACTCTGGAGAGAGAACA 3141 Left GCTGAGATTACAGGTGTGAGCTACT 3142 Right ATTAGAGCACTCTGGAGAGAGAACA 3143 Left ATTTTATTGAATTCCTTTCCAATCC 3144 Right CTCTGGAGAGAGAACAAATAAATGG 3145 Left TTATTGAATTCCTTTCCAATCCTTT 3146 Right ATTAGAGCACTCTGGAGAGAGAACA 3147 Left TTTATTGAATTCCTTTCCAATCCTT 3148 Right ATTAGAGCACTCTGGAGAGAGAACA 3149 Left TTTTATTGAATTCCTTTCCAATCCT 3150 Right ATTAGAGCACTCTGGAGAGAGAACA 3151 Left ATGCTTTCCTCCTCTATGCTATTTC 3152 Right AGCACTCTGGAGAGAGAACAAATAA 3153 Left TTTTATGCTTTCCTCCTCTATGCTA 3154 Right CTCTGGAGAGAGAACAAATAAATGG 3155 Left ATGCTTTCCTCCTCTATGCTATTTC 3156 Right ATTAGAGCACTCTGGAGAGAGAACA 3157 Left ATTCCTTTCCAATCCTTTCAGTAAC 3158 Right GAGCACTCTGGAGAGAGAACAAATA 3159 Left ATTCCTTTCCAATCCTTTCAGTAAC 3160 Right TCTGGAGAGAGAACAAATAAATGGT KIT Exon9-11 2 kb 3161 Left CCACATCCCAAGTGTTTTATGTATT 3162 Right TTCTCTATGGCAAACCTATCAAAAG 3163 Left CCACATCCCAAGTGTTTTATGTATT 3164 Right GTTCTCTATGGCAAACCTATCAAAA 3165 Left CCACATCCCAAGTGTTTTATGTATT 3166 Right ATGTTGTCCAGAGACATTTTCCTAC 3167 Left CACTAGGTCACCAAAGTGCTTATTC 3168 Right TTCTCTATGGCAAACCTATCAAAAG 3169 Left CCACATCCCAAGTGTTTTATGTATT 3170 Right CATTTTCCTACGATGTTCTCTATGG 3171 Left CCACATCCCAAGTGTTTTATGTATT 3172 Right ATGTTCTCTATGGCAAACCTATCAA 3173 Left CCACATCCCAAGTGTTTTATGTATT 3174 Right AATGTTGTCCAGAGACATTTTCCTA 3175 Left CCACATCCCAAGTGTTTTATGTATT 3176 Right AGGAATTAAAAACAATGTTGTCCAG 3177 Left TCACCAAAGTGCTTATTCTTAGACA 3178 Right TTCTCTATGGCAAACCTATCAAAAG 3179 Left TTTGTTTTAAAAGTATGCCACATCC 3180 Right ATGTTGTCCAGAGACATTTTCCTAC 3181 Left TCACCAAAGTGCTTATTCTTAGACA 3182 Right GTTCTCTATGGCAAACCTATCAAAA 3183 Left CTAGGTCACCAAAGTGCTTATTCTT 3184 Right TTCTCTATGGCAAACCTATCAAAAG 3185 Left TTTGTTTTAAAAGTATGCCACATCC 3186 Right CATTTTCCTACGATGTTCTCTATGG 3187 Left CACATCCCAAGTGTTTTATGTATT 3188 Right GGAATTAAAAACAATGTTGTCCAGA 3189 Left TTTGTTTTAAAAGTATGCCACATCC 3190 Right AATGTTGTCCAGAGACATTTTCCTA 3191 Left CTAGGTCACCAAAGTGCTTATTCTT 3192 Right GTTCTCTATGGCAAACCTATCAAAA 3193 Left AGGTCACCAAAGTGCTTATTCTTAG 3194 Right GTTCTCTATGGCAAACCTATCAAAA 3195 Left TTTGTTTTAAAAGTATGCCACATCC 3196 Right AGGAATTAAAAACAATGTTGTCCAG 3197 Left TTTGTTTTAAAAGTATGCCACATCC 3198 Right TTGTGCAGTTTCAAAATCAATAAAG 3199 Left TCACCAAAGTGCTTATTCTTAGACA 3200 Right ATGTTCTCTATGGCAAACCTATCAA KIT Exon10 130-150 bases 3201 Left TCCACATTTCTCTTCCATTGTA 3202 Right GAGAACAAATAAATGGTTAC 3203 Left CCACATTTCTCTTCCATTGTA 3204 Right AGAGAACAAATAAATGGTTAC 3205 Left TCCACATTTCTCTTCCATTGT 3206 Right GAGAACAAATAAATGGTTAC 3207 Left CCACATTTCTCTTCCATTGT 3208 Right AGAGAACAAATAAATGGTTAC 3209 Left TCCACATTTCTCTTCCATTGTA 3210 Right GAGAACAAATAAATGGTTA 3211 Left CACATTTCTCTTCCATTGTA 3212 Right GAGAGAACAAATAAATGGTTAC 3213 Left CCACATTTCTCTTCCATTGTA 3214 Right AGAGAACAAATAAATGGTTA 3215 Left CCACATTTCTCTTCCATTGTA 3216 Right GAGAACAAATAAATGGTTAC 3217 Left TCCACATTTCTCTTCCATTG 3218 Right GAGAACAAATAAATGGTTAC 3219 Left TCCACATTTCTCTTCCATTGT 3220 Right GAGAACAAATAAATGGTTA 3221 Left CCACATTTCTCTTCCATTG 3222 Right AGAGAACAAATAAATGGTTAC 3223 Left CACATTTCTCTTCCATTGT 3224 Right GAGAGAACAAATAAATGGTTAC 3225 Left CCACATTTCTCTTCCATTGT 3226 Right AGAGAACAAATAAATGGTTA 3227 Left CCACATTTCTCTTCCATTGT 3228 Right GAGAACAAATAAATGGTTAC 3229 Left CATTTCTCTTCCATTGTA 3230 Right GAGAGAGAACAAATAAATGGTTAC 3231 Left ACATTTCTCTTCCATTGTA 3232 Right AGAGAGAACAAATAAATGGTTAC 3233 Left CACATTTCTCTTCCATTGTA 3234 Right GAGAGAACAAATAAATGGTTA 3235 Left CCACATTTCTCTTCCATTGTA 3236 Right GAGAACAAATAAATGGTTA 3237 Left CACATTTCTCTTCCATTGTA 3238 Right AGAGAACAAATAAATGGTTAC 2239 Left TCCACATTTCTCTTCCATTG 2240 Right GAGAACAAATAAATGGTTA KIT Exon10 151-200 bases 3241 Left AGTTTGTGATTCCACATTTCTCTTC 3242 Right AGCACTCTGGAGAGAGAACAAATAA 3243 Left AGTTTGTGATTCCACATTTCTCTTC 3244 Right GCACTCTGGAGAGAGAACAAATAAA 3245 Left AGTTTGTGATTCCACATTTCTCTTC 3246 Right GAGCACTCTGGAGAGAGAACAAATA 3247 Left AGTTTGTGATTCCACATTTCTCTTC 3248 Right TCTGGAGAGAGAACAAATAAATGGT 3249 Left AAGTTTGTGATTCCACATTTCTCTT 3250 Right AGCACTCTGGAGAGAGAACAAATAA 3251 Left AAAGTTTGTGATTCCACATTTCTCT 3252 Right AGCACTCTGGAGAGAGAACAAATAA 3253 Left GATTCCACATTTCTCTTCCATTGTA 3254 Right AGCACTCTGGAGAGAGAACAAATAA 3255 Left AAGTTTGTGATTCCACATTTCTCTT 3256 Right GCACTCTGGAGAGAGAACAAATAAA 3257 Left AAAGTTTGTGATTCCACATTTCTCT 3258 Right GCACTCTGGAGAGAGAACAAATAAA 3259 Left AAGTTTGTGATTCCACATTTCTCTT 3260 Right GAGCACTCTGGAGAGAGAACAAATA 3261 Left AAAGTTTGTGATTCCACATTTCTCT 3262 Right GAGCACTCTGGAGAGAGAACAAATA 3263 Left CAAAGTTTGTGATTCCACATTTCTC 3264 Right AGCACTCTGGAGAGAGAACAAATAA 3265 Left CAAAGTTTGTGATTCCACATTTCTC 3266 Right GCACTCTGGAGAGAGAACAAATAAA 3267 Left CAAAGTTTGTGATTCCACATTTCTC 3268 Right CTCTGGAGAGAGAACAAATAAATGG 3269 Left AGTTTGTGATTCCACATTTCTCTTC 3270 Right CTCTGGAGAGAGAACAAATAAATGGT 3271 Left CAAAGTTTGTGATTCCACATTTCTC 3272 Right GAGCACTCTGGAGAGAGAACAAATA 3273 Left CAAAGTTTGTGATTCCACATTTCTC 3274 Right TCTGGAGAGAGAACAAATAAATGGT 3275 Left CAAAGTTTGTGATTCCACATTTCT 3276 Right AGCACTCTGGAGAGAGAACAAATAA 3277 Left GTGATTCCACATTTCTCTTCCATT 3278 Right AGCACTCTGGAGAGAGAACAAATAA 3279 Left CAAAGTTTGTGATTCCACATTTCT 3280 Right ATTAGAGCACTCTGGAGAGAGAACA KIT Exon10 201-300 bases 3281 Left GTACAATGTAACCAAGGTGAAGCTC 3282 Right AGCACTCTGGAGAGAGAACAAATAA 3283 Left GAGTACAATGTAACCAAGGTGAAGC 3284 Right AGCACTCTGGAGAGAGAACAAATAA 3285 Left TACAATGTAACCAAGGTGAAGCTCT 3286 Right AGCACTCTGGAGAGAGAACAAATAA 3287 Left TACAATGTAACCAAGGTGAAGCTCT 3288 Right ATTAGAGCACTCTGGAGAGAGAACA 3289 Left GTACAATGTAACCAAGGTGAAGCTC 3290 Right GCACTCTGGAGAGAGAACAAATAAA 3291 Left GAGTACAATGTAACCAAGGTGAAGC 3292 Right GCACTCTGGAGAGAGAACAAATAAA 3293 Left GTACAATGTAACCAAGGTGAAGCTC 3294 Right CTCTGGAGAGAGAACAAATAAATGG 3295 Left GAGTACAATGTAACCAAGGTGAAGC 3296 Right CTCTGGAGAGAGAACAAATAAATGG 3297 Left TACAATGTAACCAAGGTGAAGCTCT 3298 Right GCACTCTGGAGAGAGAACAAATAAA 3299 Left TACAATGTAACCAAGGTGAAGCTCT 3300 Right CTCTGGAGAGAGAACAAATAAATGG 3301 Left GTACAATGTAACCAAGGTGAAGCTC 3302 Right GAGCACTCTGGAGAGAGAACAAATA 3303 Left GAGTACAATGTAACCAAGGTGAAGC 3304 Right GAGCACTCTGGAGAGAGAACAAATA 3305 Left GTACAATGTAACCAAGGTGAAGCTC 3306 Right TCTGGAGAGAGAACAAATAAATGGT 3307 Left GAGTACAATGTAACCAAGGTGAAGC 3308 Right TCTGGAGAGAGAACAAATAAATGGT 3309 Left TACAATGTAACCAAGGTGAAGCTCT 3310 Right GAGCACTCTGGAGAGAGAACAAATA 3311 Left TACAATGTAACCAAGGTGAAGCTCT 3312 Right TCTGGAGAGAGAACAAATAAATGGT 3313 Left CTCTGAGACTCACATAGCTTTGCAT 3314 Right AGCACTCTGGAGAGAGAACAAATAA 3315 Left GTACAATGTAACCAAGGTGAAGCTC 3316 Right TAGAGCACTCTGGAGAGAGAACAAA 3317 Left CTCTGAGACTCACATAGCTTTGCAT 3318 Right ATTAGAGCACTCTGGAGAGAGAACA 3319 Left TACAATGTAACCAAGGTGAAGCTC 3320 Right AGCACTCTGGAGAGAGAACAAATAA KIT Exon10 301-400 bases 3321 Left TCTATTCTGCAGTATTGTGGTTTCA 3322 Right AGCACTCTGGAGAGAGAACAAATAA 3323 Left TCTGCAGTATTGTGGTTTCAAGTTA 3324 Right AGCACTCTGGAGAGAGAACAAATAA 3325 Left TCTATTCTGCAGTATTGTGGTTTCA 3326 Right ATTAGAGCACTCTGGAGAGAGAACA 3327 Left TCTGCAGTATTGTGGTTTCAAGTTA 3328 Right ATTAGAGCACTCTGGAGAGAGAACA 3329 Left TCTATTCTGCAGTATTGTGGTTTCA 3330 Right GCACTCTGGAGAGAGAACAAATAAA 3331 Left ATTCTGCAGTATTGTGGTTTCAAGT 3332 Right AGCACTCTGGAGAGAGAACAAATAA 3333 Left TCTATTCTGCAGTATTGTGGTTTCA 3334 Right CTCTGGAGAGAGAACAAATAAATGG 3335 Left TCTGCAGTATTGTGGTTTCAAGTTA 3336 Right GCACTCTGGAGAGAGAACAAATAAA 3337 Left TATTCTGCAGTATTGTGGTTTCAAG 3338 Right AGCACTCTGGAGAGAGAACAAATAA 3339 Left CTATTCTGCAGTATTGTGGTTTCAA 3340 Right AGCACTCTGGAGAGAGAACAAATAA 3341 Left TCTGCAGTATTGTGGTTTCAAGTTA 3342 Right CTCTGGAGAGAGAACAAATAAATGG 3343 Left GAGTACAATGTAACCAAGGTGAAGC 3344 Right ATTAGAGCACTCTGGAGAGAGAACA 3345 Left GTACAATGTAACCAAGGTGAAGCTC 3346 Right ATTAGAGCACTCTGGAGAGAGAACA 3347 Left ATTCTGCAGTATTGTGGTTTCAAGT 3348 Right ATTAGAGCACTCTGGAGAGAGAACA 3349 Left TATTCTGCAGTATTGTGGTTTCAAG 3350 Right ATTAGAGCACTCTGGAGAGAGAACA 3351 Left CTATTCTGCAGTATTGTGGTTTCAA 3352 Right ATTAGAGCACTCTGGAGAGAGAACA 3353 Left TCTATTCTGCAGTATTGTGGTTTCA 3354 Right TCTGGAGAGAGAACAAATAAATGGT 3355 Left TCTGCAGTATTGTGGTTTCAAGTTA 3356 Right GAGCACTCTGGAGAGAGAACAAATA 3357 Left TCTGCAGTATTGTGGTTTCAAGTTA 3358 Right TCTGGAGAGAGAACAAATAAATGGT 3359 Left ATTCTGCAGTATTGTGGTTTCAAGT 3360 Right GCACTCTGGAGAGAGAACAAATAAA KIT Exon11 151-200 bases 3361 Left AAGGTGATCTATTTTTCCCTTTCTC 3362 Right GAAAGCCCCTGTTTCATACTGAC 3363 Left AAGGTGATCTATTTTTCCCTTTCTC 3364 Right AAAGCCCCTGTTTCATACTGAC 3365 Left AAAGGTGATCTATTTTTCCCTTTCT 3366 Right GAAAGCCCCTGTTTCATACTGAC 3367 Left AAAGGTGATCTATTTTTCCCTTTCT 3368 Right AAAGCCCCTGTTTCATACTGAC 3369 Left AGGTGATCTATTTTTCCCTTTCTCC 3370 Right GAAAGCCCCTGTTTCATACTGAC 3371 Left AAGGTGATCTATTTTTCCCTTTCTC 3372 Right ATGGAAAGCCCCTGTTTCATACT 3373 Left GGTGATCTATTTTTCCCTTTCTCC 3374 Right GAAAGCCCCTGTTTCATACTGAC 3375 Left AAAGGTGATCTATTTTTCCCTTTCTC 3376 Right GAAAGCCCCTGTTTCATACTGAC 3377 Left AGGTGATCTATTTTTCCCTTTCTCC 3378 Right AAAGCCCCTGTTTCATACTGAC 3379 Left AAGGTGATCTATTTTTCCCTTTCTC 3380 Right ATGGAAAGCCCCTGTTTCATAC 3381 Left GGTGATCTATTTTTCCCTTTCTCC 3382 Right AAAGCCCCTGTTTCATACTGAC 3383 Left AAAGGTGATCTATTTTTCCCTTTCTC 3384 Right AAAGCCCCTGTTTCATACTGAC 3385 Left CTATTTTTCCCTTTCTCCCCACAG 3386 Right TTATGTGTACCCAAAAAGGTGACAT 3387 Left AAGGTGATCTATTTTTCCCTTTCTC 3388 Right GGAAAGCCCCTGTTTCATACT 3389 Left AAAGGTGATCTATTTTTCCCTTTCT 3390 Right ATGGAAAGCCCCTGTTTCATACT 3391 Left AAGGTGATCTATTTTTCCCTTTCTC 3392 Right ATGGAAAGCCCCTGTTTCATA 3393 Left TATTTTTCCCTTTCTCCCCACAG 3394 Right TTATGTGTACCCAAAAAGGTGACAT 3395 Left CTATTTTTCCCTTTCTCCCCACA 3396 Right TTATGTGTACCCAAAAAGGTGACAT 3397 Left AAAGGTGATCTATTTTTCCCTTTCT 3398 Right CAAAAAGGTGACATGGAAAGC 3399 Left AAAGGTGATCTATTTTTCCCTTTCT 3400 Right ATGGAAAGCCCCTGTTTCATAC KIT Exon11 201-300 bases 3401 Left TTATTTGTTCTCTCTCCAGAGTGCT 3402 Right TTATGTGTACCCAAAAAGGTGACAT 3403 Left TGTTCTCTCTCCAGAGTGCTCTAAT 3404 Right TTATGTGTACCCAAAAAGGTGACAT 3405 Left GTTCTCTCTCCAGAGTGCTCTAATG 3406 Right TTATGTGTACCCAAAAAGGTGACAT 3407 Left CAGAGTGCTCTAATGACTGAGACAA 3408 Right TTATGTGTACCCAAAAAGGTGACAT 3409 Left CTCTCTCCAGAGTGCTCTAATGACT 3410 Right TTATGTGTACCCAAAAAGGTGACAT 3411 Left TTATTTGTTCTCTCTCCAGAGTGCT 3412 Right TGTTATGTGTACCCAAAAAGGTGAC 3413 Left TTATTTGTTCTCTCTCCAGAGTGCT 3414 Right GTTATGTGTACCCAAAAAGGTGACA 3415 Left AAGGTGATCTATTTTTCCCTTTCTC 3416 Right TTATGTGTACCCAAAAAGGTGACAT 3417 Left AAGGTGATCTATTTTTCCCTTTCTC 3418 Right GCAATTTCACAGAAAACTCATTGTT 3419 Left TTATTTGTTCTCTCTCCAGAGTGCT 3420 Right CTGTTATGTGTACCCAAAAAGGTG 3421 Left TCTCTCTCCAGAGTGCTCTAATGAC 3422 Right TTATGTGTACCCAAAAAGGTGACAT 3423 Left TGTTCTCTCTCCAGAGTGCTCTAAT 3424 Right TGTTATGTGTACCCAAAAAGGTGAC 3425 Left GTTCTCTCTCCAGAGTGCTCTAATG 3426 Right TGTTATGTGTACCCAAAAAGGTGAC 3427 Left TGTTCTCTCTCCAGAGTGCTCTAAT 3428 Right GTTATGTGTACCCAAAAAGGTGACA 3429 Left GTTCTCTCTCCAGAGTGCTCTAATG 3430 Right GTTATGTGTACCCAAAAAGGTGACA 3431 Left GTTCTCTCTCCAGAGTGCTCTAATG 3432 Right CTGTTATGTGTACCCAAAAAGGTG 3433 Left TGTTCTCTCTCCAGAGTGCTCTAAT 3434 Right CTGTTATGTGTACCCAAAAAGGTG 3435 Left CAGAGTGCTCTAATGACTGAGACAA 3436 Right GTTATGTGTACCCAAAAAGGTGACA 3437 Left CAGAGTGCTCTAATGACTGAGACAA 3438 Right TGTTATGTGTACCCAAAAAGGTGAC 3439 Left TTATTTGTTCTCTCTCCAGAGTGCT 3440 Right CTGTTATGTGTACCCAAAAAGGTGA KIT Exon11 301-400 bases 3441 Left TTATTTGTTCTCTCTCCAGAGTGCT 3442 Right TTCTCTATGGCAAACCTATCAAAAG 3443 Left TTATTTGTTCTCTCTCCAGAGTGCT 3444 Right GTTCTCTATGGCAAACCTATCAAAA 3445 Left TGTTCTCTCTCCAGAGTGCTCTAAT 3446 Right TTCTCTATGGCAAACCTATCAAAAG 3447 Left GTTCTCTCTCCAGAGTGCTCTAATG 3448 Right TTCTCTATGGCAAACCTATCAAAAG 3449 Left CAGAGTGCTCTAATGACTGAGACAA 3450 Right TTCTCTATGGCAAACCTATCAAAAG 3451 Left GTTCTCTCTCCAGAGTGCTCTAATG 3452 Right GTTCTCTATGGCAAACCTATCAAAA 3453 Left TGTTCTCTCTCCAGAGTGCTCTAAT 3454 Right GTTCTCTATGGCAAACCTATCAAAA 3455 Left CAGAGTGCTCTAATGACTGAGACAA 3456 Right GTTCTCTATGGCAAACCTATCAAAA 3457 Left TTATTTGTTCTCTCTCCAGAGTGCT 3458 Right ATGTTGTCCAGAGACATTTTCCTAC 3459 Left TGTTCTCTCTCCAGAGTGCTCTAAT 3460 Right ATGTTGTCCAGAGACATTTTCCTAC 3461 Left GTTCTCTCTCCAGAGTGCTCTAATG 3462 Right ATGTTGTCCAGAGACATTTTCCTAC 3463 Left TTATTTGTTCTCTCTCCAGAGTGCT 3464 Right CATTTTCCTACGATGTTCTCTATGG 3465 Left CTCTCTCCAGAGTGCTCTAATGACT 3466 Right TTCTCTATGGCAAACCTATCAAAAG 3467 Left TTATTTGTTCTCTCTCCAGAGTGCT 3468 Right ATGTTCTCTATGGCAAACCTATCAA 3469 Left CAGAGTGCTCTAATGACTGAGACAA 3470 Right ATGTTGTCCAGAGACATTTTCCTAC 3471 Left TTATTTGTTCTCTCTCCAGAGTGCT 3472 Right AATGTTGTCCAGAGACATTTTCCTA 3473 Left CTCTCTCCAGAGTGCTCTAATGACT 3474 Right GTTCTCTATGGCAAACCTATCAAAA 3475 Left GTTCTCTCTCCAGAGTGCTCTAATG 3476 Right CATTTTCCTACGATGTTCTCTATGG 3477 Left TGTTCTCTCTCCAGAGTGCTCTAAT 3478 Right CATTTTCCTACGATGTTCTCTATGG 3479 Left GTTCTCTCTCCAGAGTGCTCTAATG 3480 Right ATGTTCTCTATGGCAAACCTATCAA KIT Exon12 130-150 bases 3481 Left CCTTGTTGTCTTCCTTCCTACAG 3482 Right GCAGTACCATACAGGAACTTAC 3483 Left TGTTGTCTTCCTTCCTACAG 3484 Right CATGCAGTACCATACAGGAACTTAC 3485 Left CTTGTTGTCTTCCTTCCTACAG 3486 Right TGCAGTACCATACAGGAACTTAC 3487 Left TTGTTGTCTTCCTTCCTACAG 3488 Right ATGCAGTACCATACAGGAACTTAC 3489 Left CCTTGTTGTCTTCCTTCCTACA 3490 Right GCAGTACCATACAGGAACTTAC 3491 Left CCTTGTTGTCTTCCTTCCTACAG 3492 Right GCAGTACCATACAGGAACTTA 3493 Left TGTTGTCTTCCTTCCTACAG 3494 Right CATGCAGTACCATACAGGAACTTA 3495 Left CTTGTTGTCTTCCTTCCTACAG 3496 Right TGCAGTACCATACAGGAACTTA 3497 Left CTTGTTGTCTTCCTTCCTACA 3498 Right TGCAGTACCATACAGGAACTTAC 3499 Left TTGTTGTCTTCCTTCCTACAG 3500 Right TGCAGTACCATACAGGAACTTAC 3501 Left TGTTGTCTTCCTTCCTACA 3502 Right CATGCAGTACCATACAGGAACTTAC 3503 Left TTGTTGTCTTCCTTCCTACAG 3504 Right ATGCAGTACCATACAGGAACTTA 3505 Left CCTTGTTGTCTTCCTTCCTACAG 3506 Right GCAGTACCATACAGGAACTT 3507 Left TGTTGTCTTCCTTCCTACAG 3508 Right CATGCAGTACCATACAGGAACTT 3509 Left ACCTTGTTGTCTTCCTTCCTACAG 3510 Right CAGTACCATACAGGAACTTAC 3511 Left TTGTTGTCTTCCTTCCTACA 3512 Right ATGCAGTACCATACAGGAACTTAC 3513 Left CTTGTTGTCTTCCTTCCTACAG 3514 Right TGCAGTACCATACAGGAACTT 3515 Left CCTTGTTGTCTTCCTTCCTACA 3516 Right GCAGTACCATACAGGAACTTA 3517 Left TGTTGTCTTCCTTCCTACAG 3518 Right ATGCAGTACCATACAGGAACTTAC 3519 Left TTGTTGTCTTCCTTCCTACAG 3520 Right ATGCAGTACCATACAGGAACTT KIT Exon12 151-200 bases 3521 Left TACCTTGTTGTCTTCCTTCCTACAG 3522 Right CATGCAGTACCATACAGGAACTTAC 3523 Left TTACCTTGTTGTCTTCCTTCCTACA 3524 Right CATGCAGTACCATACAGGAACTTAC 3525 Left TACCTTGTTGTCTTCCTTCCTACAG 3526 Right GCATGCAGTACCATACAGGAACTTA 3527 Left TTACCTTGTTGTCTTCCTTCCTACA 3528 Right GCATGCAGTACCATACAGGAACTTA 3529 Left ACCTTGTTGTCTTCCTTCCTACAG 3530 Right CATGCAGTACCATACAGGAACTTAC 3531 Left ACCACTTACCTTGTTGTCTTCCTTC 3532 Right CATGCAGTACCATACAGGAACTTAC 3533 Left CTTACCTTGTTGTCTTCCTTCCTAC 3534 Right CATGCAGTACCATACAGGAACTTAC 3535 Left ACTTACCTTGTTGTCTTCCTTCCTA 3536 Right CATGCAGTACCATACAGGAACTTAC 3537 Left CATCACCACTTACCTTGTTGTCTTC 3538 Right CATGCAGTACCATACAGGAACTTAC 3539 Left CACTTACCTTGTTGTCTTCCTTCCT 3540 Right CATGCAGTACCATACAGGAACTTAC 3541 Left CCACTTACCTTGTTGTCTTCCTTC 3542 Right CATGCAGTACCATACAGGAACTTAC 3543 Left CACTTACCTTGTTGTCTTCCTTCC 3544 Right CATGCAGTACCATACAGGAACTTAC 3545 Left ACTTACCTTGTTGTCTTCCTTCCTAC 3546 Right CATGCAGTACCATACAGGAACTTAC 3547 Left ACCACTTACCTTGTTGTCTTCCTT 3548 Right CATGCAGTACCATACAGGAACTTAC 3549 Left CATCACCACTTACCTTGTTGTCTT 3550 Right CATGCAGTACCATACAGGAACTTAC 3551 Left ACCTTGTTGTCTTCCTTCCTACAG 3552 Right GCATGCAGTACCATACAGGAACTTA 3553 Left ACCACTTACCTTGTTGTCTTCCTTC 3554 Right GCATGCAGTACCATACAGGAACTTA 3555 Left TACCTTGTTGTCTTCCTTCCTACA 3556 Right CATGCAGTACCATACAGGAACTTAC 3557 Left CTTACCTTGTTGTCTTCCTTCCTAC 3558 Right GCATGCAGTACCATACAGGAACTTA 3559 Left ACTTACCTTGTTGTCTTCCTTCCTA 3560 Right GCATGCAGTACCATACAGGAACTTA KIT Exon12 201-300 bases 3561 Left CTTTTGATAGGTTTGCCATAGAGAA 3562 Right CATGCAGTACCATACAGGAACTTAC 3563 Left AGAACATCGTAGGAAAATGTCTCTG 3564 Right CATGCAGTACCATACAGGAACTTAC 3565 Left CCATAGAGAACATCGTAGGAAAATG 3566 Right CATGCAGTACCATACAGGAACTTAC 3567 Left CTTTTGATAGGTTTGCCATAGAGAA 3568 Right GCATGCAGTACCATACAGGAACTTA 3569 Left TCCTTTATTGATTTTGAAACTGCAC 3570 Right CATGCAGTACCATACAGGAACTTAC 3571 Left TGATAGGTTTGCCATAGAGAACATC 3572 Right CATGCAGTACCATACAGGAACTTAC 3573 Left AGAACATCGTAGGAAAATGTCTCTG 3574 Right GCATGCAGTACCATACAGGAACTTA 3575 Left CCATAGAGAACATCGTAGGAAAATG 3576 Right GCATGCAGTACCATACAGGAACTTA 3577 Left ATTCCTTTATTGATTTTGAAACTGC 3578 Right CATGCAGTACCATACAGGAACTTAC 3579 Left ATGTCTCTGGACAACATTGTTTTTA 3580 Right CATGCAGTACCATACAGGAACTTAC 3581 Left GCCATAGAGAACATCGTAGGAAAAT 3582 Right CATGCAGTACCATACAGGAACTTAC 3583 Left TTGATAGGTTTGCCATAGAGAACA 3584 Right CATGCAGTACCATACAGGAACTTAC 3585 Left AATTCCTTTATTGATTTTGAAACTGC 3586 Right CATGCAGTACCATACAGGAACTTAC 3587 Left TTTGATAGGTTTGCCATAGAGAACA 3588 Right CATGCAGTACCATACAGGAACTTAC 3589 Left TCTCTGGACAACATTGTTTTTAATTC 3590 Right CATGCAGTACCATACAGGAACTTAC 3591 Left TAGGTTTGCCATAGAGAACATCGTA 3592 Right CATGCAGTACCATACAGGAACTTAC 3593 Left TGTCTCTGGACAACATTGTTTTTAAT 3594 Right CATGCAGTACCATACAGGAACTTAC 3595 Left AATGTCTCTGGACAACATTGTTTTTA 3596 Right CATGCAGTACCATACAGGAACTTAC 3597 Left TGGACAACATTGTTTTTAATTCCTT 3598 Right GCATTTTAGCAAAAAGCACAACT 3599 Left TCCTTTATTGATTTTGAAACTGCAC 3600 Right GCATGCAGTACCATACAGGAACTTA KIT Exon12 301-400 bases 3601 Left TGAAACAATGAGTTTTCTGTGAAAT 3602 Right CATGCAGTACCATACAGGAACTTAC 3603 Left CTGAAACAATGAGTTTTCTGTGAAAT 3604 Right CATGCAGTACCATACAGGAACTTAC 3605 Left CTTTTTGGGTACACATAACAGTGAC 3606 Right CATGCAGTACCATACAGGAACTTAC 3607 Left TTTTGGGTACACATAACAGTGACTT 3608 Right CATGCAGTACCATACAGGAACTTAC 3609 Left TTTTTGGGTACACATAACAGTGACTT 3610 Right CATGCAGTACCATACAGGAACTTAC 3611 Left CTTTTGATAGGTTTGCCATAGAGAA 3612 Right GCATTTTAGCAAAAAGCACAACT 3613 Left ACCTGAAACAATGAGTTTTCTGTGA 3614 Right CATGCAGTACCATACAGGAACTTAC 3615 Left AGAACATCGTAGGAAAATGTCTCTG 3616 Right GCATTTTAGCAAAAAGCACAACT 3617 Left AAACAATGAGTTTTCTGTGAAATTG 3618 Right CATGCAGTACCATACAGGAACTTAC 3619 Left GAAACAATGAGTTTTCTGTGAAATTG 3620 Right CATGCAGTACCATACAGGAACTTAC 3621 Left CCTTTTTGGGTACACATAACAGTGA 3622 Right CATGCAGTACCATACAGGAACTTAC 3623 Left CTTTTGATAGGTTTGCCATAGAGAA 3624 Right ATTTTAGCAAAAAGCACAACTGG 3625 Left ACCTGAAACAATGAGTTTTCTGTG 3626 Right CATGCAGTACCATACAGGAACTTAC 3627 Left AGAACATCGTAGGAAAATGTCTCTG 3628 Right ATTTTAGCAAAAAGCACAACTGG 3629 Left CCATAGAGAACATCGTAGGAAAATG 3630 Right GCATTTTAGCAAAAAGCACAACT 3631 Left TGAAACAATGAGTTTTCTGTGAAAT 3632 Right GCATGCAGTACCATACAGGAACTTA 3633 Left CCTGAAACAATGAGTTTTCTGTGA 3634 Right CATGCAGTACCATACAGGAACTTAC 3635 Left CCATAGAGAACATCGTAGGAAAATG 3636 Right ATTTTAGCAAAAAGCACAACTGG 3637 Left GTTCCACCTGAAACAATGAGTTTT 3638 Right CATGCAGTACCATACAGGAACTTAC 3639 Left CTGAAACAATGAGTTTTCTGTGAAAT 3640 Right GCATGCAGTACCATACAGGAACTTA KIT Exon13 131-150 bases 3641 Left TGCATGTTTCCAATTTTAG 3642 Right CAGCTTGGACACGGCTTTAC 3643 Left AATGCATGTTTCCAATTTTAG 3644 Right GCTTGGACACGGCTTTAC 3645 Left TGCATGTTTCCAATTTTAG 3646 Right CAGCTTGGACACGGCTTTA 3647 Left ATGCATGTTTCCAATTTTAG 3648 Right AGCTTGGACACGGCTTTAC 3649 Left TGCATGTTTCCAATTTTAG 3650 Right CAGCTTGGACACGGCTTT 3651 Left TGCATGTTTCCAATTTTAG 3652 Right AGCTTGGACACGGCTTTAC 3653 Left AATGCATGTTTCCAATTTTA 3654 Right GCTTGGACACGGCTTTAC 3655 Left ATGCATGTTTCCAATTTTAG 3656 Right AGCTTGGACACGGCTTTA 3657 Left TGCATGTTTCCAATTTTA 3658 Right CAGCTTGGACACGGCTTTAC 3659 Left ATGCATGTTTCCAATTTTAG 3660 Right GCTTGGACACGGCTTTAC 3661 Left TGCATGTTTCCAATTTTA 3662 Right CAGCTTGGACACGGCTTTA 3663 Left ATGCATGTTTCCAATTTTA 3664 Right AGCTTGGACACGGCTTTAC 3665 Left AATGCATGTTTCCAATTTT 3666 Right GCTTGGACACGGCTTTAC 3667 Left TGCATGTTTCCAATTTTA 3668 Right CAGCTTGGACACGGCTTT 3669 Left TGCATGTTTCCAATTTTAG 3670 Right AGCTTGGACACGGCTTTA 3671 Left ATGCATGTTTCCAATTTT 3672 Right AGCTTGGACACGGCTTTAC 3673 Left GCATGTTTCCAATTTTAG 3674 Right CAGCTTGGACACGGCTTTAC 3675 Left TGCATGTTTCCAATTTTAG 3676 Right GCTTGGACACGGCTTTAC 3677 Left GCATGTTTCCAATTTTAG 3678 Right CAGCTTGGACACGGCTTTA 3679 Left TGCATGTTTCCAATTTTA 3680 Right AGCTTGGACACGGCTTTAC KIT Exon13 151-200 bases 3681 Left TGCTAAAATGCATGTTTCCAAT 3682 Right CATGTTTTGATAACCTGACAGACAA 3683 Left TAAAATGCATGTTTCCAATTTTAG 3684 Right CATGTTTTGATAACCTGACAGACAA 3685 Left TGCTAAAATGCATGTTTCCAAT 3686 Right TTGATAACCTGACAGACAATAAAAGG 3687 Left TAAAATGCATGTTTCCAATTTTAG 3688 Right TTGATAACCTGACAGACAATAAAAGG 3689 Left TGCTAAAATGCATGTTTCCAAT 3690 Right TGATAACCTGACAGACAATAAAAGG 3691 Left TAAAATGCATGTTTCCAATTTTAG 3692 Right TGATAACCTGACAGACAATAAAAGG 3693 Left AAAATGCATGTTTCCAATTTTAG 3694 Right CATGTTTTGATAACCTGACAGACAA 3695 Left TGCTAAAATGCATGTTTCCAAT 3696 Right CATGTTTTGATAACCTGACAGACAAT 3697 Left TAAAATGCATGTTTCCAATTTTAG 3698 Right CATGTTTTGATAACCTGACAGACAAT 3699 Left TGCTAAAATGCATGTTTCCAAT 3700 Right ATGTTTTGATAACCTGACAGACAAT 3701 Left TGCTAAAATGCATGTTTCCAAT 3702 Right TGTTTTGATAACCTGACAGACAATAA 3703 Left TAAAATGCATGTTTCCAATTTTAG 3704 Right ATGTTTTGATAACCTGACAGACAAT 3705 Left TGCTAAAATGCATGTTTCCAAT 3706 Right CATGTTTTGATAACCTGACAGACA 3707 Left TAAAATGCATGTTTCCAATTTTAG 3708 Right TGTTTTGATAACCTGACAGACAATAA 3709 Left TAAAATGCATGTTTCCAATTTTAG 3710 Right CATGTTTTGATAACCTGACAGACA 3711 Left TGCTAAAATGCATGTTTCCAAT 3712 Right CTGACAGACAATAAAAGGCAGCTT 3713 Left TAAAATGCATGTTTCCAATTTTAG 3714 Right CTGACAGACAATAAAAGGCAGCTT 3715 Left TGCTAAAATGCATGTTTCCAAT 3716 Right TGTTTTGATAACCTGACAGACAATA 3717 Left TAAAATGCATGTTTCCAATTTTAG 3718 Right TGTTTTGATAACCTGACAGACAATA 3719 Left AAAATGCATGTTTCCAATTTTAG 3720 Right TTGATAACCTGACAGACAATAAAAGG KIT Exon13 201-300 bases 3721 Left CTTGACATCAGTTTGCCAGTTGT 3722 Right GAGAGAACAACAGTCTGGGTAAAAA 3723 Left TGCTAAAATGCATGTTTCCAAT 3724 Right GAGAGAACAACAGTCTGGGTAAAAA 3725 Left TAAAATGCATGTTTCCAATTTTAG 3726 Right GAGAGAACAACAGTCTGGGTAAAAA 3727 Left TTGACATCAGTTTGCCAGTTGT 3728 Right GAGAGAACAACAGTCTGGGTAAAAA 3729 Left CTTGACATCAGTTTGCCAGTTGT 3730 Right AGAGAGAACAACAGTCTGGGTAAAA 3731 Left TGCTAAAATGCATGTTTCCAAT 3732 Right AGAGAGAACAACAGTCTGGGTAAAA 3733 Left TGCTAAAATGCATGTTTCCAAT 3734 Right AAGAGAGAACAACAGTCTGGGTAAA 3735 Left TAAAATGCATGTTTCCAATTTTAG 3736 Right AAGAGAGAACAACAGTCTGGGTAAA 3737 Left CTTGACATCAGTTTGCCAGTTG 3738 Right GAGAGAACAACAGTCTGGGTAAAAA 3739 Left TTGACATCAGTTTGCCAGTTGT 3740 Right AGAGAGAACAACAGTCTGGGTAAAA 3741 Left TAAAATGCATGTTTCCAATTTTAG 3742 Right ACAATGAGGAAAACAAAATCTAGCA 3743 Left CTTGACATCAGTTTGCCAGTTG 3744 Right AGAGAGAACAACAGTCTGGGTAAAA 3745 Left CTTGACATCAGTTTGCCAGTTG 3746 Right AAGAGAGAACAACAGTCTGGGTAAA 3747 Left TGCTAAAATGCATGTTTCCAAT 3748 Right GGAAAACAAAATCTAGCAAGAGAGA 3749 Left TGCTAAAATGCATGTTTCCAAT 3750 Right GAGGAAAACAAAATCTAGCAAGAGA 3751 Left CTTGACATCAGTTTGCCAGTTG 3752 Right CATGTTTTGATAACCTGACAGACAA 3753 Left TAAAATGCATGTTTCCAATTTTAG 3754 Right GGAAAACAAAATCTAGCAAGAGAGA 3755 Left TAAAATGCATGTTTCCAATTTTAG 3756 Right GAGGAAAACAAAATCTAGCAAGAGA 3757 Left ATCAGTTTGCCAGTTGTGCTTT 3758 Right GAGAGAACAACAGTCTGGGTAAAAA 3759 Left ATCAGTTTGCCAGTTGTGCTT 3760 Right GAGAGAACAACAGTCTGGGTAAAAA KIT Exon13 301-400 bases 3761 Left CTTGACATCAGTTTGCCAGTTGT 3762 Right ACGACAATAACTAGGGTATGTCCTG 3763 Left TGCTAAAATGCATGTTTCCAAT 3764 Right ACGACAATAACTAGGGTATGTCCTG 3765 Left TGCTAAAATGCATGTTTCCAAT 3766 Right TCGTTGATGTTACAAATACGACAAT 3767 Left TAAAATGCATGTTTCCAATTTTAG 3768 Right ACGACAATAACTAGGGTATGTCCTG 3769 Left TAAAATGCATGTTTCCAATTTTAG 3770 Right TCGTTGATGTTACAAATACGACAAT 3771 Left CTTGACATCAGTTTGCCAGTTGT 3772 Right GCTGTTCTACCCCATAATGATAAAA 3773 Left TGCTAAAATGCATGTTTCCAAT 3774 Right GCTGATGTCGTTGATGTTACAAATA 3775 Left TGCTAAAATGCATGTTTCCAAT 3776 Right GCTGTTCTACCCCATAATGATAAAA 3777 Left TAAAATGCATGTTTCCAATTTTAG 3778 Right GCTGATGTCGTTGATGTTACAAATA 3779 Left TTGACATCAGTTTGCCAGTTGT 3780 Right ACGACAATAACTAGGGTATGTCCTG 3781 Left TTGACATCAGTTTGCCAGTTGT 3782 Right GCTGTTCTACCCCATAATGATAAAA 3783 Left CTTGACATCAGTTTGCCAGTTG 3784 Right ACGACAATAACTAGGGTATGTCCTG 3785 Left CTTGACATCAGTTTGCCAGTTGT 3786 Right GAGGAAAACAAAATCTAGCAAGAGA 3787 Left CTTGACATCAGTTTGCCAGTTG 3788 Right GCTGTTCTACCCCATAATGATAAAA 3789 Left TTGACATCAGTTTGCCAGTTGT 3790 Right GAGGAAAACAAAATCTAGCAAGAGA 3791 Left CTTGACATCAGTTTGCCAGTTG 3792 Right ACAATGAGGAAAACAAAATCTAGCA 3793 Left CTTGACATCAGTTTGCCAGTTG 3794 Right GAGGAAAACAAAATCTAGCAAGAGA 3795 Left CTTGACATCAGTTTGCCAGTTG 3796 Right GGAAAACAAAATCTAGCAAGAGAGA 3797 Left ATCAGTTTGCCAGTTGTGCTTT 3798 Right ACGACAATAACTAGGGTATGTCCTG 3799 Left ATCAGTTTGCCAGTTGTGCTTT 3800 Right GCTGTTCTACCCCATAATGATAAAA KIT Exon10-11 301-400 bases 3801 Left GATTCCACATTTCTCTTCCATTGTA 3802 Right TTATGTGTACCCAAAAAGGTGACAT 3803 Left GATTCCACATTTCTCTTCCATTGTA 3804 Right TATGTGTACCCAAAAAGGTGACAT 3805 Left GATTCCACATTTCTCTTCCATTGTA 3806 Right TTATGTGTACCCAAAAAGGTGACA 3807 Left GATTCCACATTTCTCTTCCATTGT 3808 Right TTATGTGTACCCAAAAAGGTGACAT 3809 Left ATTCCACATTTCTCTTCCATTGTA 3810 Right TTATGTGTACCCAAAAAGGTGACAT 3811 Left GATTCCACATTTCTCTTCCATTG 3812 Right TTATGTGTACCCAAAAAGGTGACAT 3813 Left GATTCCACATTTCTCTTCCATTGT 3814 Right TATGTGTACCCAAAAAGGTGACAT 3815 Left GATTCCACATTTCTCTTCCATTGTA 3816 Right ATGTGTACCCAAAAAGGTGACAT 3817 Left GATTCCACATTTCTCTTCCATTGT 3818 Right TTATGTGTACCCAAAAAGGTGACA 3819 Left GATTCCACATTTCTCTTCCATTGTA 3820 Right TATGTGTACCCAAAAAGGTGACA 3821 Left AGTTTGTGATTCCACATTTCTCTTC 3822 Right GAAAGCCCCTGTTTCATACTGAC 3823 Left ATTCCACATTTCTCTTCCATTGTA 3824 Right GTTATGTGTACCCAAAAAGGTGACA 3825 Left ATTCCACATTTCTCTTCCATTGTA 3826 Right TATGTGTACCCAAAAAGGTGACAT 3827 Left AGTTTGTGATTCCACATTTCTCTTC 3828 Right AAAGCCCCTGTTTCATACTGAC 3829 Left TGATTCCACATTTCTCTTCCATT 3830 Right TATGTGTACCCAAAAAGGTGACAT 3831 Left GATTCCACATTTCTCTTCCATTG 3832 Right TATGTGTACCCAAAAAGGTGACAT 3833 Left ATTCCACATTTCTCTTCCATTGTA 3834 Right TTATGTGTACCCAAAAAGGTGACA 3835 Left GATTCCACATTTCTCTTCCATTG 3836 Right TTATGTGTACCCAAAAAGGTGACA 3837 Left AAGTTTGTGATTCCACATTTCTCTT 3838 Right GAAAGCCCCTGTTTCATACTGAC 3839 Left AAAGTTTGTGATTCCACATTTCTCT 3840 Right GAAAGCCCCTGTTTCATACTGAC KIT Exon10-11 401-500 bases 3841 Left AGTTTGTGATTCCACATTTCTCTTC 3842 Right TTATGTGTACCCAAAAAGGTGACAT 3843 Left AGTTTGTGATTCCACATTTCTCTTC 3844 Right GCAATTTCACAGAAAACTCATTGTT 3845 Left AGTTTGTGATTCCACATTTCTCTTC 3846 Right ATTTCACAGAAAACTCATTGTTTCA 3847 Left AAGTTTGTGATTCCACATTTCTCTT 3848 Right TTATGTGTACCCAAAAAGGTGACAT 3849 Left AAAGTTTGTGATTCCACATTTCTCT 3850 Right TTATGTGTACCCAAAAAGGTGACAT 3851 Left AAGTTTGTGATTCCACATTTCTCTT 3852 Right GCAATTTCACAGAAAACTCATTGTT 3853 Left AAAGTTTGTGATTCCACATTTCTCT 3854 Right GCAATTTCACAGAAAACTCATTGTT 3855 Left AGTTTGTGATTCCACATTTCTCTTC 3856 Right GTTATGTGTACCCAAAAAGGTGACA 3857 Left AGTTTGTGATTCCACATTTCTCTTC 3858 Right TGTTATGTGTACCCAAAAAGGTGAC 3859 Left GATTCCACATTTCTCTTCCATTGTA 3860 Right TTATGTGTACCCAAAAAGGTGACAT 3861 Left GATTCCACATTTCTCTTCCATTGTA 3862 Right GCAATTTCACAGAAAACTCATTGTT 3863 Left AGTTTGTGATTCCACATTTCTCTTC 3864 Right CTGTTATGTGTACCCAAAAAGGTG 3865 Left AGTTTGTGATTCCACATTTCTCTTC 3866 Right TCACAGAAAACTCATTGTTTCAGGT 3867 Left AGTTTGTGATTCCACATTTCTCTTC 3868 Right AATTTCACAGAAAACTCATTGTTTCA 3869 Left AAGTTTGTGATTCCACATTTCTCTT 3870 Right ATTTCACAGAAAACTCATTGTTTCA 3871 Left AAAGTTTGTGATTCCACATTTCTCT 3872 Right ATTTCACAGAAAACTCATTGTTTCA 3873 Left AGTTTGTGATTCCACATTTCTCTTC 3874 Right CTGTTATGTGTACCCAAAAAGGTGA 3875 Left GATTCCACATTTCTCTTCCATTGTA 3876 Right ATTTCACAGAAAACTCATTGTTTCA 3877 Left AGTTTGTGATTCCACATTTCTCTTC 3878 Right ACAGAAAACTCATTGTTTCAGGTG 3879 Left AGTTTGTGATTCCACATTTCTCTTC 3880 Right CACAGAAAACTCATTGTTTCAGGT KIT Exon10-11 501-600 bases 3881 Left AGTTTGTGATTCCACATTTCTCTTC 3882 Right TTCTCTATGGCAAACCTATCAAAAG 3883 Left AGTTTGTGATTCCACATTTCTCTTC 3884 Right GTTCTCTATGGCAAACCTATCAAAA 3885 Left AGTTTGTGATTCCACATTTCTCTTC 3886 Right ATGTTGTCCAGAGACATTTTCCTAC 3887 Left AGTTTGTGATTCCACATTTCTCTTC 3888 Right CATTTTCCTACGATGTTCTCTATGG 3889 Left AGTTTGTGATTCCACATTTCTCTTC 3890 Right ATGTTCTCTATGGCAAACCTATCAA 3891 Left AGTTTGTGATTCCACATTTCTCTTC 3892 Right CATTTGTGCAGTTTCAAAATCAATA 3893 Left AGTTTGTGATTCCACATTTCTCTTC 3894 Right AATGTTGTCCAGAGACATTTTCCTA 3895 Left AGTTTGTGATTCCACATTTCTCTTC 3896 Right AGGAATTAAAAACAATGTTGTCCAG 3897 Left AGTTTGTGATTCCACATTTCTCTTC 3898 Right TTGTGCAGTTTCAAAATCAATAAAG 3899 Left AGTTTGTGATTCCACATTTCTCTTC 3900 Right GGAATTAAAAACAATGTTGTCCAGA 3901 Left AAGTTTGTGATTCCACATTTCTCTT 3902 Right TTCTCTATGGCAAACCTATCAAAAG 3903 Left AAAGTTTGTGATTCCACATTTCTCT 3904 Right TTCTCTATGGCAAACCTATCAAAAG 3905 Left AAGTTTGTGATTCCACATTTCTCTT 3906 Right GTTCTCTATGGCAAACCTATCAAAA 3907 Left AAAGTTTGTGATTCCACATTTCTCT 3908 Right GTTCTCTATGGCAAACCTATCAAAA 3909 Left GATTCCACATTTCTCTTCCATTGTA 3910 Right TTCTCTATGGCAAACCTATCAAAAG 3911 Left GATTCCACATTTCTCTTCCATTGTA 3912 Right GTTCTCTATGGCAAACCTATCAAAA 3913 Left AAGTTTGTGATTCCACATTTCTCTT 3914 Right ATGTTGTCCAGAGACATTTTCCTAC 3915 Left AAAGTTTGTGATTCCACATTTCTCT 3916 Right ATGTTGTCCAGAGACATTTTCCTAC 3917 Left GATTCCACATTTCTCTTCCATTGTA 3918 Right ATGTTGTCCAGAGACATTTTCCTAC 3919 Left AAGTTTGTGATTCCACATTTCTCTT 3920 Right CATTTTCCTACGATGTTCTCTATGG KIT Exon10-11 601-800 bases 3921 Left TCTATTCTGCAGTATTGTGGTTTCA 3922 Right TTCTCTATGGCAAACCTATCAAAAG 3923 Left TCTGCAGTATTGTGGTTTCAAGTTA 3924 Right TTCTCTATGGCAAACCTATCAAAAG 3925 Left TCTATTCTGCAGTATTGTGGTTTCA 3926 Right GTTCTCTATGGCAAACCTATCAAAA 3927 Left TCTGCAGTATTGTGGTTTCAAGTTA 3928 Right GTTCTCTATGGCAAACCTATCAAAA 3929 Left TCTATTCTGCAGTATTGTGGTTTCA 3930 Right ATGTTGTCCAGAGACATTTTCCTAC 3931 Left GGCAGGAATTTGATTGAAGTATAAA 3932 Right TTCTCTATGGCAAACCTATCAAAAG 3933 Left TCTGCAGTATTGTGGTTTCAAGTTA 3934 Right ATGTTGTCCAGAGACATTTTCCTAC 3935 Left GGCAGGAATTTGATTGAAGTATAAA 3936 Right GTTCTCTATGGCAAACCTATCAAAA 3937 Left TCTATTCTGCAGTATTGTGGTTTCA 3938 Right CATTTTCCTACGATGTTCTCTATGG 3939 Left TCTATTCTGCAGTATTGTGGTTTCA 3940 Right ATGTTCTCTATGGCAAACCTATCAA 3941 Left TCTATTCTGCAGTATTGTGGTTTCA 3942 Right AATGTTGTCCAGAGACATTTTCCTA 3943 Left TCTGCAGTATTGTGGTTTCAAGTTA 3944 Right CATTTTCCTACGATGTTCTCTATGG 3945 Left TCTGCAGTATTGTGGTTTCAAGTTA 3946 Right ATGTTCTCTATGGCAAACCTATCAA 3947 Left GTACAATGTAACCAAGGTGAAGCTC 3948 Right TTCTCTATGGCAAACCTATCAAAAG 3949 Left GAGTACAATGTAACCAAGGTGAAGC 3950 Right TTCTCTATGGCAAACCTATCAAAAG 3951 Left ATTCTGCAGTATTGTGGTTTCAAGT 3952 Right TTCTCTATGGCAAACCTATCAAAAG 3953 Left TCTGCAGTATTGTGGTTTCAAGTTA 3954 Right CATTTGTGCAGTTTCAAAATCAATA 3955 Left TCTGCAGTATTGTGGTTTCAAGTTA 3956 Right AATGTTGTCCAGAGACATTTTCCTA 3957 Left TCTGCAGTATTGTGGTTTCAAGTTA 3958 Right AGGAATTAAAAACAATGTTGTCCAG 3959 Left GTACAATGTAACCAAGGTGAAGCTC 3960 Right GTTCTCTATGGCAAACCTATCAAAA KIT Exon12-13 301-400 bases 3961 Left TACCTTGTTGTCTTCCTTCCTACAG 3962 Right CATGTTTTGATAACCTGACAGACAA 3963 Left TTACCTTGTTGTCTTCCTTCCTACA 3964 Right CATGTTTTGATAACCTGACAGACAA 3965 Left TACCTTGTTGTCTTCCTTCCTACAG 3966 Right TTGATAACCTGACAGACAATAAAAGG 3967 Left ACCTTGTTGTCTTCCTTCCTACAG 3968 Right CATGTTTTGATAACCTGACAGACAA 3969 Left TACCTTGTTGTCTTCCTTCCTACAG 3970 Right TGATAACCTGACAGACAATAAAAGG 3971 Left ACCACTTACCTTGTTGTCTTCCTTC 3972 Right CATGTTTTGATAACCTGACAGACAA 3973 Left CTTACCTTGTTGTCTTCCTTCCTAC 3974 Right CATGTTTTGATAACCTGACAGACAA 3975 Left ACTTACCTTGTTGTCTTCCTTCCTA 3976 Right CATGTTTTGATAACCTGACAGACAA 3977 Left CACTTACCTTGTTGTCTTCCTTCCT 3978 Right CATGTTTTGATAACCTGACAGACAA 3979 Left TTACCTTGTTGTCTTCCTTCCTACA 3980 Right TTGATAACCTGACAGACAATAAAAGG 3981 Left TACCTTGTTGTCTTCCTTCCTACAG 3982 Right CATGTTTTGATAACCTGACAGACAAT 3983 Left TTACCTTGTTGTCTTCCTTCCTACA 3984 Right TGATAACCTGACAGACAATAAAAGG 3985 Left CACTTACCTTGTTGTCTTCCTTCC 3986 Right CATGTTTTGATAACCTGACAGACAA 3987 Left CCACTTACCTTGTTGTCTTCCTTC 3988 Right CATGTTTTGATAACCTGACAGACAA 3989 Left ACTTACCTTGTTGTCTTCCTTCCTAC 3990 Right CATGTTTTGATAACCTGACAGACAA 3991 Left ACCACTTACCTTGTTGTCTTCCTT 3992 Right CATGTTTTGATAACCTGACAGACAA 3993 Left TTACCTTGTTGTCTTCCTTCCTACA 3994 Right CATGTTTTGATAACCTGACAGACAAT 3995 Left TACCTTGTTGTCTTCCTTCCTACAG 3996 Right ATGTTTTGATAACCTGACAGACAAT 3997 Left TACCTTGTTGTCTTCCTTCCTACAG 3998 Right TGTTTTGATAACCTGACAGACAATAA 3999 Left TACCTTGTTGTCTTCCTTCCTACAG 4000 Right CATGTTTTGATAACCTGACAGACA KIT Exon12-13 401-500 bases 4001 Left TACCTTGTTGTCTTCCTTCCTACAG 4002 Right GAGAGAACAACAGTCTGGGTAAAAA 4003 Left TACCTTGTTGTCTTCCTTCCTACAG 4004 Right AGAGAGAACAACAGTCTGGGTAAAA 4005 Left TACCTTGTTGTCTTCCTTCCTACAG 4006 Right AAGAGAGAACAACAGTCTGGGTAAA 4007 Left TGGACAACATTGTTTTTAATTCCTT 4008 Right CATGTTTTGATAACCTGACAGACAA 4009 Left AGAACATCGTAGGAAAATGTCTCTG 4010 Right CATGTTTTGATAACCTGACAGACAA 4011 Left TACCTTGTTGTCTTCCTTCCTACAG 4012 Right ACAATGAGGAAAACAAAATCTAGCA 4013 Left TTACCTTGTTGTCTTCCTTCCTACA 4014 Right GAGAGAACAACAGTCTGGGTAAAAA 4015 Left CCATAGAGAACATCGTAGGAAAATG 4016 Right CATGTTTTGATAACCTGACAGACAA 4017 Left CTGGACAACATTGTTTTTAATTCCT 4018 Right CATGTTTTGATAACCTGACAGACAA 4019 Left TTACCTTGTTGTCTTCCTTCCTACA 4020 Right AGAGAGAACAACAGTCTGGGTAAAA 4021 Left TTACCTTGTTGTCTTCCTTCCTACA 4022 Right AAGAGAGAACAACAGTCTGGGTAAA 4023 Left TCTGGACAACATTGTTTTTAATTCC 4024 Right CATGTTTTGATAACCTGACAGACAA 4025 Left TTACCTTGTTGTCTTCCTTCCTACA 4026 Right ACAATGAGGAAAACAAAATCTAGCA 4027 Left TGGACAACATTGTTTTTAATTCCTT 4028 Right TTATAATCTAGCATTGCCAAAATCA 4029 Left TACCTTGTTGTCTTCCTTCCTACAG 4030 Right CAATGAGGAAAACAAAATCTAGCAA 4031 Left TACCTTGTTGTCTTCCTTCCTACAG 4032 Right AACAATGAGGAAAACAAAATCTAGC 4033 Left TACCTTGTTGTCTTCCTTCCTACAG 4034 Right TTATAATCTAGCATTGCCAAAATCA 4035 Left TCCTTTATTGATTTTGAAACTGCAC 4036 Right CATGTTTTGATAACCTGACAGACAA 4037 Left TGATAGGTTTGCCATAGAGAACATC 4038 Right CATGTTTTGATAACCTGACAGACAA 4039 Left TGGACAACATTGTTTTTAATTCCTT 4040 Right CAGTTTATAATCTAGCATTGCCAAAA KIT Exon12-13 501-600 bases 4041 Left TACCTTGTTGTCTTCCTTCCTACAG 4042 Right ATGAGATATTCAAGAGGCTGATGTC 4043 Left TACCTTGTTGTCTTCCTTCCTACAG 4044 Right GATGAGATATTCAAGAGGCTGATGT 4045 Left CTTTTGATAGGTTTGCCATAGAGAA 4046 Right GAGAGAACAACAGTCTGGGTAAAAA 4047 Left TGGACAACATTGTTTTTAATTCCTT 4048 Right GAGAGAACAACAGTCTGGGTAAAAA 4049 Left AGAACATCGTAGGAAAATGTCTCTG 4050 Right GAGAGAACAACAGTCTGGGTAAAAA 4051 Left CTTTTGATAGGTTTGCCATAGAGAA 4052 Right AGAGAGAACAACAGTCTGGGTAAAA 4053 Left CTTTTGATAGGTTTGCCATAGAGAA 4054 Right AAGAGAGAACAACAGTCTGGGTAAA 4055 Left TGGACAACATTGTTTTTAATTCCTT 4056 Right AGAGAGAACAACAGTCTGGGTAAAA 4057 Left TGGACAACATTGTTTTTAATTCCTT 4058 Right AAGAGAGAACAACAGTCTGGGTAAA 4059 Left AGAACATCGTAGGAAAATGTCTCTG 4060 Right AGAGAGAACAACAGTCTGGGTAAAA 4061 Left AGAACATCGTAGGAAAATGTCTCTG 4062 Right AAGAGAGAACAACAGTCTGGGTAAA 4063 Left CCATAGAGAACATCGTAGGAAAATG 4064 Right GAGAGAACAACAGTCTGGGTAAAAA 4065 Left TACCTTGTTGTCTTCCTTCCTACAG 4066 Right ACGACAATAACTAGGGTATGTCCTG 4067 Left CTTTTGATAGGTTTGCCATAGAGAA 4068 Right CATGTTTTGATAACCTGACAGACAA 4069 Left CTGGACAACATTGTTTTTAATTCCT 4070 Right GAGAGAACAACAGTCTGGGTAAAAA 4071 Left TACCTTGTTGTCTTCCTTCCTACAG 4072 Right TCGTTGATGTTACAAATACGACAAT 4073 Left TACCTTGTTGTCTTCCTTCCTACAG 4074 Right GCTGATGTCGTTGATGTTACAAATA 4075 Left TACCTTGTTGTCTTCCTTCCTACAG 4076 Right GCTGTTCTACCCCATAATGATAAAA 4077 Left TGGACAACATTGTTTTTAATTCCTT 4078 Right ACAATGAGGAAAACAAAATCTAGCA 4079 Left TTACCTTGTTGTCTTCCTTCCTACA 4080 Right ATGAGATATTCAAGAGGCTGATGTC KIT Exon12-13 601-800 bases 4081 Left CTTTTGATAGGTTTGCCATAGAGAA 4082 Right GATGAGATATTCAAGAGGCTGATGT 4083 Left CTTTTGATAGGTTTGCCATAGAGAA 4084 Right ATGAGATATTCAAGAGGCTGATGTC 4085 Left TGGACAACATTGTTTTTAATTCCTT 4086 Right TTTCAGTGGCTACATATGATCAAGA 4087 Left TGGACAACATTGTTTTTAATTCCTT 4088 Right TTCAGTGGCTACATATGATCAAGAA 4089 Left AGAACATCGTAGGAAAATGTCTCTG 4090 Right ATGAGATATTCAAGAGGCTGATGTC 4091 Left AGAACATCGTAGGAAAATGTCTCTG 4092 Right GATGAGATATTCAAGAGGCTGATGT 4093 Left AGAACATCGTAGGAAAATGTCTCTG 4094 Right TCAGTGGCTACATATGATCAAGAAA 4095 Left AGAACATCGTAGGAAAATGTCTCTG 4096 Right TTCAGTGGCTACATATGATCAAGAA 4097 Left AGAACATCGTAGGAAAATGTCTCTG 4098 Right TTTCAGTGGCTACATATGATCAAGA 4099 Left TACCTTGTTGTCTTCCTTCCTACAG 4100 Right TCAGTGGCTACATATGATCAAGAAA 4101 Left TACCTTGTTGTCTTCCTTCCTACAG 4102 Right TTCAGTGGCTACATATGATCAAGAA 4103 Left TACCTTGTTGTCTTCCTTCCTACAG 4104 Right TTTCAGTGGCTACATATGATCAAGA 4105 Left ACAGTGACTTTAAGGAACTCCAGTG 4106 Right GATGAGATATTCAAGAGGCTGATGT 4107 Left ACAGTGACTTTAAGGAACTCCAGTG 4108 Right ATGAGATATTCAAGAGGCTGATGTC 4109 Left TACCTTGTTGTCTTCCTTCCTACAG 4110 Right ATGTTTGCCTCATTTGGTGTATATT 4111 Left ACAGTGACTTTAAGGAACTCCAGTG 4112 Right GAGAGAACAACAGTCTGGGTAAAAA 4113 Left CCATAGAGAACATCGTAGGAAAATG 4114 Right GATGAGATATTCAAGAGGCTGATGT 4115 Left CCATAGAGAACATCGTAGGAAAATG 4116 Right ATGAGATATTCAAGAGGCTGATGTC 4117 Left TACCTTGTTGTCTTCCTTCCTACAG 4118 Right AAGGCTTCAAATTGGAAACTTATTT 4119 Left CTTTTGATAGGTTTGCCATAGAGAA 4120 Right ACGACAATAACTAGGGTATGTCCTG KIT Exon10-13 801-1000 bases 4121 Left AAGTTTGTGATTCCACATTTCTCTT 4122 Right CATGTTTTGATAACCTGACAGACAA 4123 Left GATTCCACATTTCTCTTCCATTGTA 4124 Right CATGTTTTGATAACCTGACAGACAA 4125 Left AGTTTGTGATTCCACATTTCTCTTC 4126 Right TTGATAACCTGACAGACAATAAAAGG 4127 Left AGTTTGTGATTCCACATTTCTCTTC 4128 Right TGATAACCTGACAGACAATAAAAGG 4129 Left CAAAGTTTGTGATTCCACATTTCTC 4130 Right CATGTTTTGATAACCTGACAGACAA 4131 Left AGTTTGTGATTCCACATTTCTCTTC 4132 Right CATGTTTTGATAACCTGACAGACAAT 4133 Left AAGTTTGTGATTCCACATTTCTCTT 4134 Right TTGATAACCTGACAGACAATAAAAGG 4135 Left AAAGTTTGTGATTCCACATTTCTCT 4136 Right TTGATAACCTGACAGACAATAAAAGG 4137 Left AAGTTTGTGATTCCACATTTCTCTT 4138 Right TGATAACCTGACAGACAATAAAAGG 4139 Left AAAGTTTGTGATTCCACATTTCTCT 4140 Right TGATAACCTGACAGACAATAAAAGG 4141 Left CAAAGTTTGTGATTCCACATTTCT 4142 Right CATGTTTTGATAACCTGACAGACAA 4143 Left GTGATTCCACATTTCTCTTCCATT 4144 Right CATGTTTTGATAACCTGACAGACAA 4145 Left AAAGTTTGTGATTCCACATTTCTCTT 4146 Right CATGTTTTGATAACCTGACAGACAA 4147 Left AGTTTGTGATTCCACATTTCTCTTC 4148 Right ATGTTTTGATAACCTGACAGACAAT 4149 Left AGTTTGTGATTCCACATTTCTCTTC 4150 Right TGTTTTGATAACCTGACAGACAATAA 4151 Left AGTTTGTGATTCCACATTTCTCTTC 4152 Right CATGTTTTGATAACCTGACAGACA 4153 Left AAGTTTGTGATTCCACATTTCTCTT 4154 Right CATGTTTTGATAACCTGACAGACAAT 4155 Left AGTTTGTGATTCCACATTTCTCTTC 4156 Right CTGACAGACAATAAAAGGCAGCTT 4157 Left AGTTTGTGATTCCACATTTCTCTTC 4158 Right TGTTTTGATAACCTGACAGACAATA 4159 Left CAAAGTTTGTGATTCCACATTTCTC 4160 Right TTGATAACCTGACAGACAATAAAAGG KIT Exon10-13 2 kb 4161 Left AATATAAACCGTCCATAAAGGAAGC 4162 Right ATTATGGAAATAATGAAGGCACAAA 4163 Left AATATAAACCGTCCATAAAGGAAGC 4164 Right TTCTGAGCGCTACTAGTTGAAAAAT 4165 Left AATATAAACCGTCCATAAAGGAAGC 4166 Right ATTTATGGGGATTATTGGAAGACAT 4167 Left TCTATTCTGCAGTATTGTGGTTTCA 4168 Right TTCTGAGCGCTACTAGTTGAAAAAT 4169 Left AATATAAACCGTCCATAAAGGAAGC 4170 Right CTGGAAAGTGAGTGAAAACCTAAAA 4171 Left AATATAAACCGTCCATAAAGGAAGC 4172 Right GACTGGAAAGTGAGTGAAAACCTAA 4173 Left AGTTTGTGATTCCACATTTCTCTTC 4174 Right GATTAATTCTGCCTCCCATAAAAAT 4175 Left ACTGCCTTTATGAAGTCATGTTAGC 4176 Right ATGAGATATTCAAGAGGCTGATGTC 4177 Left TCTGCAGTATTGTGGTTTCAAGTTA 4178 Right TTCTGAGCGCTACTAGTTGAAAAAT 4179 Left AGTTTGTGATTCCACATTTCTCTTC 4180 Right AATATTCAGCTTCAGTGGATCAGAC 4181 Left AGTTTGTGATTCCACATTTCTCTTC 4182 Right GAATCCTCTGGTTCACTTCTGTTTA 4183 Left TCTATTCTGCAGTATTGTGGTTTCA 4184 Right GAATCCTCTGGTTCACTTCTGTTTA 4185 Left AGAATCTTCCATGTTTTTCAGACAG 4186 Right GACTGGAAAGTGAGTGAAAACCTAA 4187 Left AATATAAACCGTCCATAAAGGAAGC 4188 Right ACAAGACTGGAAAGTGAGTGAAAAC 4189 Left TTTTAACACTTTGCCAGACACTGTA 4190 Right GATGAGATATTCAAGAGGCTGATGT 4191 Left TTTTAACACTTTGCCAGACACTGTA 4192 Right ATGAGATATTCAAGAGGCTGATGTC 4193 Left GGATGTTTAGGCTCTGTCTACCATA 4194 Right GAGAGAACAACAGTCTGGGTAAAAA 4195 Left TCTGCAGTATTGTGGTTTCAAGTTA 4196 Right GAATCCTCTGGTTCACTTCTGTTTA 4197 Left AATATAAACCGTCCATAAAGGAAGC 4198 Right TATGGAAATAATGAAGGCACAAACT 4199 Left AATACATGCATCACACCATACTGTC 4200 Right GAGAGAACAACAGTCTGGGTAAAAA KIT Exon10-13 5 kb 4201 Left GGATGTTTAGGCTCTGTCTACCATA 4202 Right CAGAAACTAGGTGTCCTTTTTGTGT 4203 Left GGATGTTTAGGCTCTGTCTACCATA 4204 Right ACCAATAATTTGGATGATTTTCTGA 4205 Left GGATGTTTAGGCTCTGTCTACCATA 4206 Right TAATTTGGATGATTTTCTGAACCAT 4207 Left GGATGTTTAGGCTCTGTCTACCATA 4208 Right TCTGATTAGCATAGAAACACCATGA 4209 Left GGATGTTTAGGCTCTGTCTACCATA 4210 Right GGTATCCCGTATTACCTCAAACTCT 4211 Left GGATGTTTAGGCTCTGTCTACCATA 4212 Right ACTTCCTGCTTCTGATTAGCATAGA 4213 Left ACTGCCTTTATGAAGTCATGTTAGC 4214 Right CAGAAACTAGGTGTCCTTTTTGTGT 4215 Left AATACATGCATCACACCATACTGTC 4216 Right TAATTTGGATGATTTTCTGAACCAT 4217 Left AATACATGCATCACACCATACTGTC 4218 Right ACCAATAATTTGGATGATTTTCTGA 4219 Left AATACATGCATCACACCATACTGTC 4220 Right TCTGATTAGCATAGAAACACCATGA 4221 Left AATACATGCATCACACCATACTGTC 4222 Right GGTATCCCGTATTACCTCAAACTCT 4223 Left AATACATGCATCACACCATACTGTC 4224 Right ACTTCCTGCTTCTGATTAGCATAGA 4225 Left ACTGCCTTTATGAAGTCATGTTAGC 4226 Right GGTATCCCGTATTACCTCAAACTCT 4227 Left AGAATCTTCCATGTTTTTCAGACAG 4228 Right CTCATCCCCTTTGCTACATAATAGA 4229 Left TTTTAACACTTTGCCAGACACTGTA 4230 Right CAGAAACTAGGTGTCCTTTTTGTGT 4231 Left GGATGTTTAGGCTCTGTCTACCATA 4232 Right CTCATCCCCTTTGCTACATAATAGA 4233 Left TTTTAACACTTTGCCAGACACTGTA 4234 Right GGTATCCCGTATTACCTCAAACTCT 4235 Left AATACATGCATCACACCATACTGTC 4236 Right CTCATCCCCTTTGCTACATAATAGA 4237 Left AGAATCTTCCATGTTTTTCAGACAG 4238 Right CATAATAGACTGGACCAATTTGAGG 4239 Left TTTAATCCAATTTAAGGGGATGTTT 4240 Right CAGAAACTAGGTGTCCTTTTTGTGT KIT Exon17 151-200 bases 4241 Left GTTTTCTTTTCTCCTCCAACCTAAT 4242 Right ACTGTCAAGCAGAGAATGGGTACT 4243 Left GTTTTCTTTTCTCCTCCAACCTAAT 4244 Right CTGTCAAGCAGAGAATGGGTACT 4245 Left TGAATTTAAATGGTTTTCTTTTCTCC 4246 Right ACTGTCAAGCAGAGAATGGGTACT 4247 Left GTTTTCTTTTCTCCTCCAACCTAAT 4248 Right GACTGTCAAGCAGAGAATGGGTACT 4249 Left TTTTCTTTTCTCCTCCAACCTAATA 4250 Right ACTGTCAAGCAGAGAATGGGTACT 4251 Left TAAATGGTTTTCTTTTCTCCTCCA 4252 Right ACTGTCAAGCAGAGAATGGGTACT 4253 Left TAAATGGTTTTCTTTTCTCCTCCAA 4254 Right ACTGTCAAGCAGAGAATGGGTACT 4255 Left TTAAATGGTTTTCTTTTCTCCTCCA 4256 Right ACTGTCAAGCAGAGAATGGGTACT 4257 Left GTTTTCTTTTCTCCTCCAACCTAATA 4258 Right ACTGTCAAGCAGAGAATGGGTACT 4259 Left GTTTTCTTTTCTCCTCCAACCTAAT 4260 Right GACTGTCAAGCAGAGAATGGGTA 4261 Left TGAATTTAAATGGTTTTCTTTTCTCC 4262 Right CTGTCAAGCAGAGAATGGGTACT 4263 Left TGAATTTAAATGGTTTTCTTTTCTCC 4264 Right GACTGTCAAGCAGAGAATGGGTACT 4265 Left TTTTCTTTTCTCCTCCAACCTAATA 4266 Right CTGTCAAGCAGAGAATGGGTACT 4267 Left GTTTTCTTTTCTCCTCCAACCTAA 4268 Right ACTGTCAAGCAGAGAATGGGTACT 4269 Left TTTTCTTTTCTCCTCCAACCTAATA 4270 Right GACTGTCAAGCAGAGAATGGGTACT 4271 Left TAAATGGTTTTCTTTTCTCCTCCA 4272 Right CTGTCAAGCAGAGAATGGGTACT 4273 Left TAAATGGTTTTCTTTTCTCCTCCA 4274 Right GACTGTCAAGCAGAGAATGGGTACT 4275 Left GTTTTCTTTTCTCCTCCAACCTAAT 4276 Right CAGGACTGTCAAGCAGAGAATG 4277 Left TAAATGGTTTTCTTTTCTCCTCCAA 4278 Right CTGTCAAGCAGAGAATGGGTACT 4279 Left TTAAATGGTTTTCTTTTCTCCTCCA 4280 Right CTGTCAAGCAGAGAATGGGTACT KIT Exon17 201-300 4281 Left GTTTTCTTTTCTCCTCCAACCTAAT 4282 Right ATCACAGGAAACAATTTTTATCGAA 4283 Left GTTTTCTTTTCTCCTCCAACCTAAT 4284 Right AAATGTGTGATATCCCTAGACAGGA 4285 Left GTTTTCTTTTCTCCTCCAACCTAAT 4286 Right TGTGTGATATCCCTAGACAGGATTT 4287 Left GTTTTCTTTTCTCCTCCAACCTAAT 4288 Right CACAGGAAACAATTTTTATCGAAAG 4289 Left GTTTTCTTTTCTCCTCCAACCTAAT 4290 Right AAAATGTGTGATATCCCTAGACAGG 4291 Left TGAATTTAAATGGTTTTCTTTTCTCC 4292 Right TGTGTGATATCCCTAGACAGGATTT 4293 Left TGAATTTAAATGGTTTTCTTTTCTCC 4294 Right AAATGTGTGATATCCCTAGACAGGA 4295 Left TTTTCTTTTCTCCTCCAACCTAATA 4296 Right ATCACAGGAAACAATTTTTATCGAA 4297 Left GTTTTCTTTTCTCCTCCAACCTAAT 4298 Right ATGTGTGATATCCCTAGACAGGATT 4299 Left GTTTTCTTTTCTCCTCCAACCTAAT 4300 Right AATGTGTGATATCCCTAGACAGGAT 4301 Left TTTTCTTTTCTCCTCCAACCTAATA 4302 Right AAATGTGTGATATCCCTAGACAGGA 4303 Left TTTTCTTTTCTCCTCCAACCTAATA 4304 Right TGTGTGATATCCCTAGACAGGATTT 4305 Left TGAATTTAAATGGTTTTCTTTTCTCC 4306 Right CACAGGAAACAATTTTTATCGAAAG 4307 Left TGAATTTAAATGGTTTTCTTTTCTCC 4308 Right AAAATGTGTGATATCCCTAGACAGG 4309 Left TTTTCTTTTCTCCTCCAACCTAATA 4310 Right CACAGGAAACAATTTTTATCGAAAG 4311 Left TAAATGGTTTTCTTTTCTCCTCCA 4312 Right ATCACAGGAAACAATTTTTATCGAA 4313 Left TAAATGGTTTTCTTTTCTCCTCCA 4314 Right AAATGTGTGATATCCCTAGACAGGA 4315 Left TAAATGGTTTTCTTTTCTCCTCCA 4316 Right TGTGTGATATCCCTAGACAGGATTT 4317 Left TTTTCTTTTCTCCTCCAACCTAATA 4318 Right AAAATGTGTGATATCCCTAGACAGG 4319 Left TAAATGGTTTTCTTTTCTCCTCCAA 4320 Right ATCACAGGAAACAATTTTTATCGAA KIT Exon17 301-400 4321 Left ATTCAAGGCGTACTTTTGATTTTTA 4322 Right ATCACAGGAAACAATTTTTATCGAA 4323 Left TCAAGGCGTACTTTTGATTTTTATT 4324 Right ATCACAGGAAACAATTTTTATCGAA 4325 Left TTCAAGGCGTACTTTTGATTTTTAT 4326 Right ATCACAGGAAACAATTTTTATCGAA 4327 Left ATTCAAGGCGTACTTTTGATTTTTA 4328 Right CACAGGAAACAATTTTTATCGAAAG 4329 Left GTTTTCTTTTCTCCTCCAACCTAAT 4330 Right TAGTAATGTTCAGCATACCATGCAA 4331 Left ATTCAAGGCGTACTTTTGATTTTTAT 4332 Right ATCACAGGAAACAATTTTTATCGAA 4333 Left ATCATTCAAGGCGTACTTTTGATTT 4334 Right ATCACAGGAAACAATTTTTATCGAA 4335 Left TTCAAGGCGTACTTTTGATTTTTAT 4336 Right TCGAAAGTTGAAACTAAAAATCCTTT 4337 Left ATTCAAGGCGTACTTTTGATTTTTA 4338 Right TCACAGGAAACAATTTTTATCGAA 4339 Left TCAAGGCGTACTTTTGATTTTTATT 4340 Right TCACAGGAAACAATTTTTATCGAA 4341 Left TTCAAGGCGTACTTTTGATTTTTAT 4342 Right TCACAGGAAACAATTTTTATCGAA 4343 Left ATTCAAGGCGTACTTTTGATTTTT 4344 Right ATCACAGGAAACAATTTTTATCGAA 4345 Left TTCAAGGCGTACTTTTGATTTTTATT 4346 Right ATCACAGGAAACAATTTTTATCGAA 4347 Left AGTCCTGAGAAGAAAACAGCATTTA 4348 Right ACTGTCAAGCAGAGAATGGGTACT 4349 Left ATTCAAGGCGTACTTTTGATTTTT 4350 Right CACAGGAAACAATTTTTATCGAAAG 4351 Left GTTTTCTTTTCTCCTCCAACCTAAT 4352 Right GTAATGTTCAGCATACCATGCAAAT 4353 Left CATTCAAGGCGTACTTTTGATTTT 4354 Right ATCACAGGAAACAATTTTTATCGAA 4355 Left TTCAAGGCGTACTTTTGATTTTTA 4356 Right ATCACAGGAAACAATTTTTATCGAA 4357 Left GAACATCATTCAAGGCGTACTTTT 4358 Right ATCACAGGAAACAATTTTTATCGAA 4359 Left CATTCAAGGCGTACTTTTGATTTTT 4360 Right ATCACAGGAAACAATTTTTATCGAA KIT Exon17 401-500 bases 4361 Left ATGTATTTCCCTATGAATGAAAGCA 4362 Right ATCACAGGAAACAATTTTTATCGAA 4363 Left TATTTCCCTATGAATGAAAGCAGTC 4364 Right ATCACAGGAAACAATTTTTATCGAA 4365 Left AGTCCTGAGAAGAAAACAGCATTTA 4366 Right ATCACAGGAAACAATTTTTATCGAA 4367 Left AGTCCTGAGAAGAAAACAGCATTTA 4368 Right TGTGTGATATCCCTAGACAGGATTT 4369 Left ATGTATTTCCCTATGAATGAAAGCA 4370 Right CACAGGAAACAATTTTTATCGAAAG 4371 Left TATTTCCCTATGAATGAAAGCAGTC 4372 Right CACAGGAAACAATTTTTATCGAAAG 4373 Left AGTCCTGAGAAGAAAACAGCATTTA 4374 Right CACAGGAAACAATTTTTATCGAAAG 4375 Left TCCTGAGAAGAAAACAGCATTTATT 4376 Right ATCACAGGAAACAATTTTTATCGAA 4377 Left TCCTGAGAAGAAAACAGCATTTATT 4378 Right TGTGTGATATCCCTAGACAGGATTT 4379 Left AGTCCTGAGAAGAAAACAGCATTTA 4380 Right AAAATGTGTGATATCCCTAGACAGG 4381 Left GTTTTCTTTTCTCCTCCAACCTAAT 4382 Right AATCAAGTTCATTGCTATTCTCAGG 4383 Left TCCTGAGAAGAAAACAGCATTTATT 4384 Right CACAGGAAACAATTTTTATCGAAAG 4385 Left GTTTTCTTTTCTCCTCCAACCTAAT 4386 Right GCAAAATCAAGTTCATTGCTATTCT 4387 Left GTTTTCTTTTCTCCTCCAACCTAAT 4388 Right AGCAAAATCAAGTTCATTGCTATTC 4389 Left TCCTGAGAAGAAAACAGCATTTATT 4390 Right AAAATGTGTGATATCCCTAGACAGG 4391 Left TCAAGGCGTACTTTTGATTTTTATT 4392 Right AAATGTGTGATATCCCTAGACAGGA 4393 Left TCAAGGCGTACTTTTGATTTTTATT 4394 Right TGTGTGATATCCCTAGACAGGATTT 4395 Left TTCAAGGCGTACTTTTGATTTTTAT 4396 Right AAATGTGTGATATCCCTAGACAGGA 4397 Left ATTCAAGGCGTACTTTTGATTTTTA 4398 Right AAATGTGTGATATCCCTAGACAGGA 4399 Left TTCAAGGCGTACTTTTGATTTTTAT 4400 Right TGTGTGATATCCCTAGACAGGATTT KIT Exon17 501-600 4401 Left ATGTATTTCCCTATGAATGAAAGCA 4402 Right AAATGTGTGATATCCCTAGACAGGA 4403 Left ATGTATTTCCCTATGAATGAAAGCA 4404 Right TGTGTGATATCCCTAGACAGGATTT 4405 Left TATTTCCCTATGAATGAAAGCAGTC 4406 Right AAATGTGTGATATCCCTAGACAGGA 4407 Left TATTTCCCTATGAATGAAAGCAGTC 4408 Right TGTGTGATATCCCTAGACAGGATTT 4409 Left GATTGAATTTGCAAAGGCATATTAG 4410 Right ATCACAGGAAACAATTTTTATCGAA 4411 Left ATGTATTTCCCTATGAATGAAAGCA 4412 Right AAAATGTGTGATATCCCTAGACAGG 4413 Left TATTTCCCTATGAATGAAAGCAGTC 4414 Right AAAATGTGTGATATCCCTAGACAGG 4415 Left GATTGAATTTGCAAAGGCATATTAG 4416 Right CACAGGAAACAATTTTTATCGAAAG 4417 Left AAAGGCATATTAGGAACTCTGTGAA 4418 Right ATCACAGGAAACAATTTTTATCGAA 4419 Left AAGGCATATTAGGAACTCTGTGAAA 4420 Right ATCACAGGAAACAATTTTTATCGAA 4421 Left TCAAGGCGTACTTTTGATTTTTATT 4422 Right AATCAAGTTCATTGCTATTCTCAGG 4423 Left TTCAAGGCGTACTTTTGATTTTTAT 4424 Right AATCAAGTTCATTGCTATTCTCAGG 4425 Left ATTCAAGGCGTACTTTTGATTTTTA 4426 Right AATCAAGTTCATTGCTATTCTCAGG 4427 Left TCAAGGCGTACTTTTGATTTTTATT 4428 Right GCAAAATCAAGTTCATTGCTATTCT 4429 Left TTCAAGGCGTACTTTTGATTTTTAT 4430 Right GCAAAATCAAGTTCATTGCTATTCT 4431 Left ATTCAAGGCGTACTTTTGATTTTTA 4432 Right GCAAAATCAAGTTCATTGCTATTCT 4433 Left TCAAGGCGTACTTTTGATTTTTATT 4434 Right AGCAAAATCAAGTTCATTGCTATTC 4435 Left TTCAAGGCGTACTTTTGATTTTTAT 4436 Right AGCAAAATCAAGTTCATTGCTATTC 4437 Left ATTCAAGGCGTACTTTTGATTTTTA 4438 Right AGCAAAATCAAGTTCATTGCTATTC 4439 Left AAAGGCATATTAGGAACTCTGTGAA 4440 Right TGTGTGATATCCCTAGACAGGATTT KIT Exon17 601-800 4441 Left TATTTCCCTATGAATGAAAGCAGTC 4442 Right AATCAAGTTCATTGCTATTCTCAGG 4443 Left TATTTCCCTATGAATGAAAGCAGTC 4444 Right GCAAAATCAAGTTCATTGCTATTCT 4445 Left ACATTTCCCAACAATTACCAAACTA 4446 Right ATCACAGGAAACAATTTTTATCGAA 4447 Left ACATTTCCCAACAATTACCAAACTA 4448 Right AAATGTGTGATATCCCTAGACAGGA 4449 Left ACATTTCCCAACAATTACCAAACTA 4450 Right TGTGTGATATCCCTAGACAGGATTT 4451 Left GAAGGTTAGGAATGGAAAGAATGAT 4452 Right ATCACAGGAAACAATTTTTATCGAA 4453 Left GATTGAATTTGCAAAGGCATATTAG 4454 Right AATCAAGTTCATTGCTATTCTCAGG 4455 Left TTCCCAACAATTACCAAACTAAGAA 4456 Right ATCACAGGAAACAATTTTTATCGAA 4457 Left TCCCAACAATTACCAAACTAAGAAA 4458 Right ATCACAGGAAACAATTTTTATCGAA 4459 Left TTTCCCAACAATTACCAAACTAAGA 4460 Right ATCACAGGAAACAATTTTTATCGAA 4461 Left GATTGAATTTGCAAAGGCATATTAG 4462 Right GCAAAATCAAGTTCATTGCTATTCT 4463 Left GATTGAATTTGCAAAGGCATATTAG 4464 Right AGCAAAATCAAGTTCATTGCTATTC 4465 Left TTACCAGTCCTACCCTTAAATGTCA 4466 Right ATCACAGGAAACAATTTTTATCGAA 4467 Left TCCCAACAATTACCAAACTAAGAAA 4468 Right AAATGTGTGATATCCCTAGACAGGA 4469 Left TTCCCAACAATTACCAAACTAAGAA 4470 Right AAATGTGTGATATCCCTAGACAGGA 4471 Left TTTCCCAACAATTACCAAACTAAGA 4472 Right AAATGTGTGATATCCCTAGACAGGA 4473 Left TTCCCAACAATTACCAAACTAAGAA 4474 Right TGTGTGATATCCCTAGACAGGATTT 4475 Left TTTCCCAACAATTACCAAACTAAGA 4476 Right TGTGTGATATCCCTAGACAGGATTT 4477 Left TCCCAACAATTACCAAACTAAGAAA 4478 Right TGTGTGATATCCCTAGACAGGATTT 4479 Left ATTAACCGAACAGAATGAGTTACCA 4480 Right ATCACAGGAAACAATTTTTATCGAA KIT Exon17 801-1000 bases 4481 Left ACATTTCCCAACAATTACCAAACTA 4482 Right AATCAAGTTCATTGCTATTCTCAGG 4483 Left ACATTTCCCAACAATTACCAAACTA 4484 Right GCAAAATCAAGTTCATTGCTATTCT 4485 Left ACATTTCCCAACAATTACCAAACTA 4486 Right AGCAAAATCAAGTTCATTGCTATTC 4487 Left ATGTATTTCCCTATGAATGAAAGCA 4488 Right CATGCCTTAGTTTCTCCAACTTTTA 4489 Left TATTTCCCTATGAATGAAAGCAGTC 4490 Right CATGCCTTAGTTTCTCCAACTTTTA 4491 Left TCCCAACAATTACCAAACTAAGAAA 4492 Right AATCAAGTTCATTGCTATTCTCAGG 4493 Left TTTCCCAACAATTACCAAACTAAGA 4494 Right AATCAAGTTCATTGCTATTCTCAGG 4495 Left TCCCAACAATTACCAAACTAAGAAA 4496 Right GCAAAATCAAGTTCATTGCTATTCT 4497 Left TCCCAACAATTACCAAACTAAGAAA 4498 Right AGCAAAATCAAGTTCATTGCTATTC 4499 Left TTCCCAACAATTACCAAACTAAGAA 4500 Right AGCAAAATCAAGTTCATTGCTATTC 4501 Left TTTCCCAACAATTACCAAACTAAGA 4502 Right GCAAAATCAAGTTCATTGCTATTCT 4503 Left TTTCCCAACAATTACCAAACTAAGA 4504 Right AGCAAAATCAAGTTCATTGCTATTC 4505 Left GGGTGAAGCATAGACTTGAGTTTTA 4506 Right ATCACAGGAAACAATTTTTATCGAA 4507 Left ATTAACCGAACAGAATGAGTTACCA 4508 Right AATCAAGTTCATTGCTATTCTCAGG 4509 Left TTACCAGTCCTACCCTTAAATGTCA 4510 Right GCAAAATCAAGTTCATTGCTATTCT 4511 Left TTACCAGTCCTACCCTTAAATGTCA 4512 Right AGCAAAATCAAGTTCATTGCTATTC 4513 Left GGGTGAAGCATAGACTTGAGTTTTA 4514 Right AAATGTGTGATATCCCTAGACAGGA 4515 Left GGGTGAAGCATAGACTTGAGTTTTA 4516 Right TGTGTGATATCCCTAGACAGGATTT 4517 Left AGTCCTGAGAAGAAAACAGCATTTA 4518 Right CATGCCTTAGTTTCTCCAACTTTTA 4519 Left ATTAACCGAACAGAATGAGTTACCA 4520 Right GCAAAATCAAGTTCATTGCTATTCT KIT Exon17 2 kb 4521 Left AACCAAAAGCAGAGGAAATTTAGTT 4522 Right AACACTCTTACAAAACCAAATCGAG 4523 Left AGCCATAGTTAAAATGCAGAATGTC 4524 Right AACACTCTTACAAAACCAAATCGAG 4525 Left GAGCCATAGTTAAAATGCAGAATGT 4526 Right AACACTCTTACAAAACCAAATCGAG 4527 Left AGAGCCATAGTTAAAATGCAGAATG 4528 Right AACACTCTTACAAAACCAAATCGAG 4529 Left ACATTTCCCAACAATTACCAAACTA 4530 Right AGTATGTAACCACCGTCACGATTAT 4531 Left GGGTGAAGCATAGACTTGAGTTTTA 4532 Right AGTATGTAACCACCGTCACGATTAT 4533 Left GGGTGAAGCATAGACTTGAGTTTTA 4534 Right TTGGACAACTCTTGACAAAATTACA 4535 Left GAAGGTTAGGAATGGAAAGAATGAT 4536 Right AGTATGTAACCACCGTCACGATTAT 4537 Left GGGTGAAGCATAGACTTGAGTTTTA 4538 Right ATTCAGAGGTATTGGACAACTCTTG 4539 Left GAAGGTTAGGAATGGAAAGAATGAT 4540 Right TTGGACAACTCTTGACAAAATTACA 4541 Left ATGTATTTCCCTATGAATGAAAGCA 4542 Right AAAACCCACAATTACTTTTACACCA 4543 Left TATTTCCCTATGAATGAAAGCAGTC 4544 Right AAAACCCACAATTACTTTTACACCA 4545 Left AACCAAAAGCAGAGGAAATTTAGTT 4546 Right ATTACATTATCATAAGGGGCACAAA 4547 Left GAAGGTTAGGAATGGAAAGAATGAT 4548 Right ATTCAGAGGTATTGGACAACTCTTG 4549 Left AGCCATAGTTAAAATGCAGAATGTC 4550 Right GGCAGAGAATATTATAAAGGGCAAT 4551 Left GAGCCATAGTTAAAATGCAGAATGT 4552 Right GGCAGAGAATATTATAAAGGGCAAT 4553 Left AGAGCCATAGTTAAAATGCAGAATG 4554 Right GGCAGAGAATATTATAAAGGGCAAT 4555 Left AGCCATAGTTAAAATGCAGAATGTC 4556 Right ATTACATTATCATAAGGGGCACAAA 4557 Left GAGCCATAGTTAAAATGCAGAATGT 4558 Right ATTACATTATCATAAGGGGCACAAA 4559 Left AGAGCCATAGTTAAAATGCAGAATG 4560 Right ATTACATTATCATAAGGGGCACAAA KIT Exon17 5 kb 4561 Left ATGGTTCAGAAAATCATCCAAATTA 4562 Right TCTGCCATAAAAAGCTAAATCAATC 4563 Left TCAGAAAATCATCCAAATTATTGGT 4564 Right TCTGCCATAAAAAGCTAAATCAATC 4565 Left GTATATTGCTGCAGTTGTGTGGTAG 4566 Right TAAGGGCTCCTAACCTGAGATCTAT 4567 Left TCATGGTGTTTCTATGCTAATCAGA 4568 Right TCTGCCATAAAAAGCTAAATCAATC 4569 Left GTACCTACCTATCAAGCAACCAAGA 4570 Right CTTGAGTACCATCTCACAAAAACCT 4571 Left GAGTACCTACCTATCAAGCAACCAA 4572 Right CTTGAGTACCATCTCACAAAAACCT 4573 Left TCTATGCTAATCAGAAGCAGGAAGT 4574 Right TCTGCCATAAAAAGCTAAATCAATC 4575 Left ATGGTTCAGAAAATCATCCAAATTA 4576 Right GGCCACTAAGTTGTAAGTGCTGTAT 4577 Left TCAGAAAATCATCCAAATTATTGGT 4578 Right GGCCACTAAGTTGTAAGTGCTGTAT 4579 Left GTATATTGCTGCAGTTGTGTGGTAG 4580 Right AAAACCCACAATTACTTTTACACCA 4581 Left GTACCTACCTATCAAGCAACCAAGA 4582 Right GGCCACTAAGTTGTAAGTGCTGTAT 4583 Left GAGTACCTACCTATCAAGCAACCAA 4584 Right GGCCACTAAGTTGTAAGTGCTGTAT 4585 Left TCATGGTGTTTCTATGCTAATCAGA 4586 Right TAAGGGCTCCTAACCTGAGATCTAT 4587 Left GTAACCCAGCCTAGGATTGTTAAAT 4588 Right CTTGAGTACCATCTCACAAAAACCT 4589 Left TCATGGTGTTTCTATGCTAATCAGA 4590 Right GGCCACTAAGTTGTAAGTGCTGTAT 4591 Left AACCAAAAGCAGAGGAAATTTAGTT 4592 Right CAGTGTGTCATAAAGAATCCAAGTG 4593 Left TCTATGCTAATCAGAAGCAGGAAGT 4594 Right TAAGGGCTCCTAACCTGAGATCTAT 4595 Left ATGTTTTTGTGCCTGAGTATCTTTC 4596 Right CAGTGTGTCATAAAGAATCCAAGTG 4597 Left TCTATGCTAATCAGAAGCAGGAAGT 4598 Right GGCCACTAAGTTGTAAGTGCTGTAT 4599 Left AACCAAAAGCAGAGGAAATTTAGTT 4600 Right CAATTTGCAACCTAAGATTAGGAGA

TABLE 10 KRAS Capture Primer List for NGS Panel Seq. ID Primer Sequence KRAS Exon1 169-300 bases 4601 Left CTCGGAGCTCGATTTTCCTA 4602 Right GGGACCCCTAATTCATTCACTC 4603 Left CTCGGAGCTCGATTTTCCTA 4604 Right GGGACCCCTAATTCATTCACT 4605 Left CTCGGAGCTCGATTTTCCTA 4606 Right GGACCCCTAATTCATTCACTCG 4607 Left CTCGGAGCTCGATTTTCCT 4608 Right GGGACCCCTAATTCATTCACTC 4609 Left GTGCTCGGAGCTCGATTTT 4610 Right GGGACCCCTAATTCATTCACTC 4611 Left CTCGGAGCTCGATTTTCCTA 4612 Right GGGGACCCCTAATTCATTCACT 4613 Left GCTCGGAGCTCGATTTTCCTA 4614 Right GGGACCCCTAATTCATTCACTC 4615 Left CTCGGAGCTCGATTTTCCTA 4616 Right GACCCCTAATTCATTCACTCG 4617 Left CTCGGAGCTCGATTTTCCTA 4618 Right GGGGACCCCTAATTCATTCAC 4619 Left GTGCTCGGAGCTCGATTTTC 4620 Right GGGACCCCTAATTCATTCACTC 4621 Left TGCTCGGAGCTCGATTTT 4622 Right GGGACCCCTAATTCATTCACTC 4623 Left CTCGGAGCTCGATTTTCCT 4624 Right GGGACCCCTAATTCATTCACT 4625 Left CTCGGAGCTCGATTTTCCTA 4626 Right GGGGACCCCTAATTCATTCA 4627 Left GTGCTCGGAGCTCGATTTT 4628 Right GGGACCCCTAATTCATTCACT 4629 Left TCGGAGCTCGATTTTCCTA 4630 Right GGGACCCCTAATTCATTCACTC 4631 Left CTCGGAGCTCGATTTTCCT 4632 Right GGACCCCTAATTCATTCACTCG 4633 Left GTGCTCGGAGCTCGATTTT 4634 Right GGACCCCTAATTCATTCACTCG 4635 Left GTGTGCTCGGAGCTCGATT 4636 Right GGGACCCCTAATTCATTCACTC 4637 Left CTCGGAGCTCGATTTTCCTA 4638 Right GGGGACCCCTAATTCATTCACTC 4639 Left TGCTCGGAGCTCGATTTTC 4640 Right GGGACCCCTAATTCATTCACTC KRAS Exon1 301-400 bases 4641 Left CCCGTCTGAAGAAGAATCGAG 4642 Right GGGACCCCTAATTCATTCACTC 4643 Left GTACGCCCGTCTGAAGAAGA 4644 Right GGGACCCCTAATTCATTCACTC 4645 Left CATCGATAGCTCTGCCCTCT 4646 Right GGGACCCCTAATTCATTCACTC 4647 Left ATCGATAGCTCTGCCCTCTG 4648 Right GGGACCCCTAATTCATTCACTC 4649 Left GTACGCCCGTCTGAAGAAGAATC 4650 Right GGGACCCCTAATTCATTCACTC 4651 Left GTACGCCCGTCTGAAGAAGAA 4652 Right GGGACCCCTAATTCATTCACTC 4653 Left GTACGCCCGTCTGAAGAAGAAT 4654 Right GGGACCCCTAATTCATTCACT 4655 Left TACGCCCGTCTGAAGAAGAATC 4656 Right GGGACCCCTAATTCATTCACTC 4657 Left GTACGCCCGTCTGAAGAAGAAT 4658 Right GGACCCCTAATTCATTCACTCG 4659 Left TACGCCCGTCTGAAGAAGAA 4660 Right GGGACCCCTAATTCATTCACTC 4661 Left GGAACGCATCGATAGCTCTG 4662 Right GGGACCCCTAATTCATTCACTC 4663 Left TACGCCCGTCTGAAGAAGAAT 4664 Right GGGACCCCTAATTCATTCACT 4665 Left TACGCCCGTCTGAAGAAGAAT 4666 Right GGACCCCTAATTCATTCACTCG 4667 Left CCCGTCTGAAGAAGAATCGAG 4668 Right GGGACCCCTAATTCATTCACT 4669 Left CATCGATAGCTCTGCCCTCTG 4670 Right GGGACCCCTAATTCATTCACTC 4671 Left GTACGCCCGTCTGAAGAAGA 4672 Right GGGACCCCTAATTCATTCACT 4673 Left GTACGCCCGTCTGAAGAAGAAT 4674 Right GGGGACCCCTAATTCATTCACT 4675 Left ATCGATAGCTCTGCCCTCTG 4676 Right GGGACCCCTAATTCATTCACT 4677 Left CATCGATAGCTCTGCCCTCT 4678 Right GGGACCCCTAATTCATTCACT 4679 Left GTACGCCCGTCTGAAGAAGAATC 4680 Right GGGACCCCTAATTCATTCACT KRAS Exon1 401-600 bases 4681 Left CTCTTCCCTCTTCCCACACC 4682 Right GGGACCCCTAATTCATTCACTC 4683 Left CCTCTTCCCTCTTCCCACAC 4684 Right GGGACCCCTAATTCATTCACTC 4685 Left TCTTCCCTCTTCCCACACC 4686 Right GGGACCCCTAATTCATTCACTC 4687 Left CCTCTTCCCTCTTCCCACA 4688 Right GGGACCCCTAATTCATTCACTC 4689 Left CTCTTCCCTCTTCCCACACC 4690 Right GGGACCCCTAATTCATTCACT 4691 Left CCTCTTCCCTCTTCCCACAC 4692 Right GGGACCCCTAATTCATTCACT 4693 Left TCTTCCCTCTTCCCACACC 4694 Right GGGACCCCTAATTCATTCACT 4695 Left CTCTTCCCTCTTCCCACACC 4696 Right GGACCCCTAATTCATTCACTCG 4697 Left CCTCTTCCCTCTTCCCACAC 4698 Right GGACCCCTAATTCATTCACTCG 4699 Left TCTTCCCTCTTCCCACACC 4700 Right GGACCCCTAATTCATTCACTCG 4701 Left CCTCTTCCCTCTTCCCACA 4702 Right GGGACCCCTAATTCATTCACT 4703 Left CCTCTTCCCTCTTCCCACA 4704 Right GGACCCCTAATTCATTCACTCG 4705 Left GTACGCCCGTCTGAAGAAGAAT 4706 Right GAGGAGGAAGGAAGGGGTTC 4707 Left GTACGCCCGTCTGAAGAAGAAT 4708 Right AGGAGGAAGGAAGGGGTTC 4709 Left TACGCCCGTCTGAAGAAGAAT 4710 Right GAGGAGGAAGGAAGGGGTTC 4711 Left CTCTTCCCTCTTCCCACACC 4712 Right GGGGACCCCTAATTCATTCACT 4713 Left CCTCTTCCCTCTTCCCACAC 4714 Right GGGGACCCCTAATTCATTCACT 4715 Left TCTTCCCTCTTCCCACACC 4716 Right GGGGACCCCTAATTCATTCACT 4717 Left CTCTTCCCTCTTCCCACACC 4718 Right GACCCCTAATTCATTCACTCG 4719 Left CCTCTTCCCTCTTCCCACAC 4720 Right GACCCCTAATTCATTCACTCG KRAS Exon1 601-800 bases 4721 Left GTACGCCCGTCTGAAGAAGAAT 4722 Right GTTTATACCTTCGTCCTAGAGATGC 4723 Left GTACGCCCGTCTGAAGAAGAAT 4724 Right AGTTTATACCTTCGTCCTAGAGATGC 4725 Left TACGCCCGTCTGAAGAAGAAT 4726 Right GTTTATACCTTCGTCCTAGAGATGC 4727 Left CCCGTCTGAAGAAGAATCGAG 4728 Right GTTTATACCTTCGTCCTAGAGATGC 4729 Left TACGCCCGTCTGAAGAAGAAT 4730 Right AGTTTATACCTTCGTCCTAGAGATGC 4731 Left GTACGCCCGTCTGAAGAAGA 4732 Right GTTTATACCTTCGTCCTAGAGATGC 4733 Left ATCGATAGCTCTGCCCTCTG 4734 Right GTTTATACCTTCGTCCTAGAGATGC 4735 Left CATCGATAGCTCTGCCCTCT 4736 Right GTTTATACCTTCGTCCTAGAGATGC 4737 Left GTACGCCCGTCTGAAGAAGAATC 4738 Right GTTTATACCTTCGTCCTAGAGATGC 4739 Left CCCGTCTGAAGAAGAATCGAG 4740 Right AGTTTATACCTTCGTCCTAGAGATGC 4741 Left CTCGGAGCTCGATTTTCCTA 4742 Right CTTACTCACACATCCCCTACACAC 4743 Left CTCGGAGCTCGATTTTCCTA 4744 Right GTTTATACCTTCGTCCTAGAGATGC 4745 Left CCCGTCTGAAGAAGAATCGAG 4746 Right GGAGAAGTTTATACCTTCGTCCTAGA 4747 Left GTACGCCCGTCTGAAGAAGA 4748 Right AGTTTATACCTTCGTCCTAGAGATGC 4749 Left ATCGATAGCTCTGCCCTCTG 4750 Right AGTTTATACCTTCGTCCTAGAGATGC 4751 Left CATCGATAGCTCTGCCCTCT 4752 Right AGTTTATACCTTCGTCCTAGAGATGC 4753 Left GTACGCCCGTCTGAAGAAGAATC 4754 Right AGTTTATACCTTCGTCCTAGAGATGC 4755 Left CTCGGAGCTCGATTTTCCTA 4756 Right AGTTTATACCTTCGTCCTAGAGATGC 4757 Left ATCGATAGCTCTGCCCTCTG 4758 Right GGAGAAGTTTATACCTTCGTCCTAGA 4759 Left CATCGATAGCTCTGCCCTCT 4760 Right GGAGAAGTTTATACCTTCGTCCTAGA KRAS Exon1 801-1000 bases 4761 Left ACTTTTGGTGACTGCTTGTTTATTT 4762 Right GGGACCCCTAATTCATTCACTC 4763 Left CACTTTTGGTGACTGCTTGTTTATT 4764 Right GGGACCCCTAATTCATTCACTC 4765 Left GTACGCCCGTCTGAAGAAGAAT 4766 Right ACAGTGGTCTCTAAGCACTTTCCTA 4767 Left GTACGCCCGTCTGAAGAAGAAT 4768 Right AAACACAATAACCTCAAACAGTGGT 4769 Left CTTTTGGTGACTGCTTGTTTATTTA 4770 Right GGGACCCCTAATTCATTCACTC 4771 Left GTACGCCCGTCTGAAGAAGAAT 4772 Right TAACCTCAAACAGTGGTCTCTAAGC 4773 Left GTACGCCCGTCTGAAGAAGAAT 4774 Right ACACAATAACCTCAAACAGTGGTCT 4775 Left TTTTGGTGACTGCTTGTTTATTTAC 4776 Right GGGACCCCTAATTCATTCACTC 4777 Left TACGCCCGTCTGAAGAAGAAT 4778 Right ACAGTGGTCTCTAAGCACTTTCCTA 4779 Left TACGCCCGTCTGAAGAAGAAT 4780 Right AAACACAATAACCTCAAACAGTGGT 4781 Left CACTTTTGGTGACTGCTTGTTTAT 4782 Right GGGACCCCTAATTCATTCACTC 4783 Left ACTTTTGGTGACTGCTTGTTTATTT 4784 Right GGGACCCCTAATTCATTCACT 4785 Left GTACGCCCGTCTGAAGAAGAAT 4786 Right ACAATAACCTCAAACAGTGGTCTCT 4787 Left CTTTTGGTGACTGCTTGTTTATTTAC 4788 Right GGGACCCCTAATTCATTCACTC 4789 Left ACTTTTGGTGACTGCTTGTTTATTTA 4790 Right GGGACCCCTAATTCATTCACTC 4791 Left ACTTTTGGTGACTGCTTGTTTATTT 4792 Right GGACCCCTAATTCATTCACTCG 4793 Left TACGCCCGTCTGAAGAAGAAT 4794 Right TAACCTCAAACAGTGGTCTCTAAGC 4795 Left CCCGTCTGAAGAAGAATCGAG 4796 Right ACAGTGGTCTCTAAGCACTTTCCTA 4797 Left TACGCCCGTCTGAAGAAGAAT 4798 Right ACACAATAACCTCAAACAGTGGTCT 4799 Left CCCGTCTGAAGAAGAATCGAG 4800 Right AAACACAATAACCTCAAACAGTGGT KRAS Exon1 2 kb 4801 Left GGGATTTAAATTCAGCTTTATTGGT 4802 Right ACAGTGGTCTCTAAGCACTTTCCTA 4803 Left GGGATTTAAATTCAGCTTTATTGGT 4804 Right CATCTGGGATTAACTTTTTCCTTTT 4805 Left TCAAGACTCTCCCAAGATACATTTC 4806 Right TTTGCTATTGCTGTCTACACTCAAC 4807 Left TCAAGACTCTCCCAAGATACATTTC 4808 Right ACAGTGGTCTCTAAGCACTTTCCTA 4809 Left GGGATTTAAATTCAGCTTTATTGGT 4810 Right AAACACAATAACCTCAAACAGTGGT 4811 Left TCAAGACTCTCCCAAGATACATTTC 4812 Right CATCTGGGATTAACTTTTTCCTTTT 4813 Left TCAAGACTCTCCCAAGATACATTTC 4814 Right AAACACAATAACCTCAAACAGTGGT 4815 Left GTAGAAAGGAAAGGATGACAGTTGA 4816 Right AAACACAATAACCTCAAACAGTGGT 4817 Left GATTACAGCCCGTGTAAGAGTAGAA 4818 Right ACAGTGGTCTCTAAGCACTTTCCTA 4819 Left GATTACAGCCCGTGTAAGAGTAGAA 4820 Right AAACACAATAACCTCAAACAGTGGT 4821 Left GGGATTTAAATTCAGCTTTATTGGT 4822 Right TAACCTCAAACAGTGGTCTCTAAGC 4823 Left TCAAGACTCTCCCAAGATACATTTC 4824 Right ATAAGAAATAGGGGAAAGGACAAGA 4825 Left TCAAGACTCTCCCAAGATACATTTC 4826 Right TAACCTCAAACAGTGGTCTCTAAGC 4827 Left TTCAGCTTTATTGGTGGTTTATGAT 4828 Right ACAGTGGTCTCTAAGCACTTTCCTA 4829 Left ATTCAGCTTTATTGGTGGTTTATGA 4830 Right ACAGTGGTCTCTAAGCACTTTCCTA 4831 Left GGGATTTAAATTCAGCTTTATTGGT 4832 Right ACACAATAACCTCAAACAGTGGTCT 4833 Left TTCAGCTTTATTGGTGGTTTATGAT 4834 Right CATCTGGGATTAACTTTTTCCTTTT 4835 Left ATTCAGCTTTATTGGTGGTTTATGA 4836 Right CATCTGGGATTAACTTTTTCCTTTT 4837 Left ATTCAGCTTTATTGGTGGTTTATGA 4838 Right AAACACAATAACCTCAAACAGTGGT 4839 Left TTCAGCTTTATTGGTGGTTTATGAT 4840 Right AAACACAATAACCTCAAACAGTGGT KRAS Exon1 5 kb 4841 Left GGGATTTAAATTCAGCTTTATTGGT 4842 Right CAAAGCAATTAGGAATAGATGAGGA 4843 Left ACGTAAGTAAGGAAGGGAGAACAGT 4844 Right CAAAGCAATTAGGAATAGATGAGGA 4845 Left AGCAGTAAATGAAACAGACCAAAAC 4846 Right CAAAGCAATTAGGAATAGATGAGGA 4847 Left GGGATTTAAATTCAGCTTTATTGGT 4848 Right TCCTTTCCCTCATGTAACACATAAT 4849 Left TGCTTTGAATGTTAGTCACAGAGAG 4850 Right TGATGGATCTCAAGATTTAAGAAGG 4851 Left ACGTAAGTAAGGAAGGGAGAACAGT 4852 Right AACAGTTCTCAAAATGTGGTCTAGG 4853 Left ACGTAAGTAAGGAAGGGAGAACAGT 4854 Right TCCTTTCCCTCATGTAACACATAAT 4855 Left CGAATCATGAGCCTAGATGATAACT 4856 Right ATCCAACAATTTTGTAATGGAAGAA 4857 Left CGAATCATGAGCCTAGATGATAACT 4858 Right AATCCAACAATTTTGTAATGGAAGA 4859 Left CGAATCATGAGCCTAGATGATAACT 4860 Right GAATCCAACAATTTTGTAATGGAAG 4861 Left AGCAGTAAATGAAACAGACCAAAAC 4862 Right AACAGTTCTCAAAATGTGGTCTAGG 4863 Left AGCTCTGGAGAAAAAGTAGGAAAAG 4864 Right CAAAGCAATTAGGAATAGATGAGGA 4865 Left TCAAGACTCTCCCAAGATACATTTC 4866 Right TCCTTTCCCTCATGTAACACATAAT 4867 Left TAGAGTGACTATGATCCGACATGAA 4868 Right CAAAGCAATTAGGAATAGATGAGGA 4869 Left CTCCTTTTAAAAACACTTTGGAACA 4870 Right CAAAGCAATTAGGAATAGATGAGGA 4871 Left ACGTAAGTAAGGAAGGGAGAACAGT 4872 Right TCATGTTACCAAGTAATGGGCTTAT 4873 Left GAAGAGGGTAGGGGATATCAAATAA 4874 Right CAAAGCAATTAGGAATAGATGAGGA 4875 Left GCTGGAAATTTAGCAGTAAATGAAA 4876 Right CAAAGCAATTAGGAATAGATGAGGA 4877 Left GGAAACAAGAACTTATCATGCACTT 4878 Right ATCCAACAATTTTGTAATGGAAGAA 4879 Left GGAAACAAGAACTTATCATGCACTT 4880 Right GAATCCAACAATTTTGTAATGGAAG KRAS Exon2 169-210 bases 4881 Left CATGTTCTAATATAGTCACATTTTCA 4882 Right AAGAATGGTCCTGCACCAGTAATA 4883 Left CATGTTCTAATATAGTCACATTTTCA 4884 Right AGAATGGTCCTGCACCAGTAATA 4885 Left ACATGTTCTAATATAGTCACATTTTCA 4886 Right AGAATGGTCCTGCACCAGTAATA 4887 Left GACATGTTCTAATATAGTCACATTTT CA 4888 Right GAATGGTCCTGCACCAGTAATATG 4889 Left CATGTTCTAATATAGTCACATTTTCATT 4890 Right AAGAATGGTCCTGCACCAGTAATA 4891 Left CATGTTCTAATATAGTCACATTTTCATT 4892 Right AGAATGGTCCTGCACCAGTAATA 4893 Left CATGTTCTAATATAGTCACATTTTCAT 4894 Right AAGAATGGTCCTGCACCAGTAATA 4895 Left CATGTTCTAATATAGTCACATTTTCAT 4896 Right AGAATGGTCCTGCACCAGTAATA 4897 Left CATGTTCTAATATAGTCACATTTTCA 4898 Right GAATGGTCCTGCACCAGTAATATG 4899 Left ACATGTTCTAATATAGTCACATTTTC AT 4900 Right AGAATGGTCCTGCACCAGTAATA 4901 Left GACATGTTCTAATATAGTCACATTTT CATT 4902 Right GAATGGTCCTGCACCAGTAATATG 4901 Left GACATGTTCTAATATAGTCACATTTT CATT 4902 Right GAATGGTCCTGCACCAGTAATATG 4903 Left ACATGTTCTAATATAGTCACATTTTCA 4904 Right GAATGGTCCTGCACCAGTAATATG 4905 Left GACATGTTCTAATATAGTCACATTTT CAT 4906 Right GAATGGTCCTGCACCAGTAATATG 4907 Left CATGTTCTAATATAGTCACATTTTCATT 4908 Right GAATGGTCCTGCACCAGTAATATG 4909 Left CATGTTCTAATATAGTCACATTTTCA 4910 Right AGAATGGTCCTGCACCAGTAAT 4911 Left TGACATGTTCTAATATAGTCACATTT TCAT 4912 Right ATGGTCCTGCACCAGTAATATG 4913 Left ACATGTTCTAATATAGTCACATTTTCA 4914 Right AGAATGGTCCTGCACCAGTAAT 4915 Left CATGTTCTAATATAGTCACATTTTCAT 4916 Right GAATGGTCCTGCACCAGTAATATG 4917 Left TGACATGTTCTAATATAGTCACATTT TCAT 4918 Right ATGGTCCTGCACCAGTAATATGC 4919 Left TGACATGTTCTAATATAGTCACATTT TCAT 4920 Right AATGGTCCTGCACCAGTAATATGC KRAS Exon2 211-300 bases 4921 Left AAAAGGTACTGGTGGAGTATTTGAT 4922 Right ATGAAAATGGTCAGAGAAACCTTTA 4923 Left TTAAAAGGTACTGGTGGAGTATTTGA 4924 Right ATGAAAATGGTCAGAGAAACCTTTA 4925 Left TAAAAGGTACTGGTGGAGTATTTGA 4926 Right ATGAAAATGGTCAGAGAAACCTTTA 4927 Left GGTACTGGTGGAGTATTTGATAGTG 4928 Right ATGAAAATGGTCAGAGAAACCTTTA 4929 Left AGGTACTGGTGGAGTATTTGATAGTG 4930 Right ATGAAAATGGTCAGAGAAACCTTTA 4931 Left TTTGTATTAAAAGGTACTGGTGGAG 4932 Right ATGAAAATGGTCAGAGAAACCTTTA 4933 Left AAGGTACTGGTGGAGTATTTGATAGTG 4934 Right ATGAAAATGGTCAGAGAAACCTTTA 4935 Left GGTACTGGTGGAGTATTTGATAGTGT 4936 Right ATGAAAATGGTCAGAGAAACCTTTA 4937 Left AAAAGGTACTGGTGGAGTATTTGAT 4938 Right TACTCATGAAAATGGTCAGAGAAAC 4939 Left GTTTGTATTAAAAGGTACTGGTGGA 4940 Right ATGAAAATGGTCAGAGAAACCTTTA 4941 Left TTAAAAGGTACTGGTGGAGTATTTGA 4942 Right TACTCATGAAAATGGTCAGAGAAAC 4943 Left AGGTACTGGTGGAGTATTTGATAGTGT 4944 Right ATGAAAATGGTCAGAGAAACCTTTA 4945 Left ATTAAAAGGTACTGGTGGAGTATTTGA 4946 Right ATGAAAATGGTCAGAGAAACCTTTA 4947 Left TTTGTATTAAAAGGTACTGGTGGAGT 4948 Right ATGAAAATGGTCAGAGAAACCTTTA 4949 Left GTTTGTATTAAAAGGTACTGGTGGAG 4950 Right ATGAAAATGGTCAGAGAAACCTTTA 4951 Left AAAAGGTACTGGTGGAGTATTTGATA 4952 Right ATGAAAATGGTCAGAGAAACCTTTA 4953 Left TAAAAGGTACTGGTGGAGTATTTGA 4954 Right TACTCATGAAAATGGTCAGAGAAAC 4955 Left TTGTATTAAAAGGTACTGGTGGAGT 4956 Right ATGAAAATGGTCAGAGAAACCTTTA 4957 Left GGTACTGGTGGAGTATTTGATAGTG 4958 Right TACTCATGAAAATGGTCAGAGAAAC 4959 Left AAAGGTACTGGTGGAGTATTTGATA 4960 Right ATGAAAATGGTCAGAGAAACCTTTA KRAS Exon2 301-400 bases 4961 Left AAAAGGTACTGGTGGAGTATTTGAT 4962 Right CCAAGGAAAGTAAAGTTCCCATATT 4963 Left TTAAAAGGTACTGGTGGAGTATTTGA 4964 Right CCAAGGAAAGTAAAGTTCCCATATT 4965 Left TAAAAGGTACTGGTGGAGTATTTGA 4966 Right CCAAGGAAAGTAAAGTTCCCATATT 4967 Left AAAAGGTACTGGTGGAGTATTTGAT 4968 Right TAACTTGAAACCCAAGGTACATTTC 4969 Left AAAAGGTACTGGTGGAGTATTTGAT 4970 Right GAAACCCAAGGTACATTTCAGATAA 4971 Left TTAAAAGGTACTGGTGGAGTATTTGA 4972 Right TAACTTGAAACCCAAGGTACATTTC 4973 Left GGTACTGGTGGAGTATTTGATAGTG 4974 Right CCAAGGAAAGTAAAGTTCCCATATT 4975 Left TGAAGTACAGTTCATTACGATACACG 4976 Right ATGAAAATGGTCAGAGAAACCTTTA 4977 Left TAAAAGGTACTGGTGGAGTATTTGA 4978 Right TAACTTGAAACCCAAGGTACATTTC 4979 Left AGGTACTGGTGGAGTATTTGATAGTG 4980 Right CCAAGGAAAGTAAAGTTCCCATATT 4981 Left CAGTCAACTGGAATTTTCATGATT 4982 Right ATGAAAATGGTCAGAGAAACCTTTA 4983 Left TTAAAAGGTACTGGTGGAGTATTTG 4984 Right CCAAGGAAAGTAAAGTTCCCATATT 4985 Left AAGGTACTGGTGGAGTATTTGATAGTG 4986 Right CCAAGGAAAGTAAAGTTCCCATATT 4987 Left GGTACTGGTGGAGTATTTGATAGTGT 4988 Right CCAAGGAAAGTAAAGTTCCCATATT 4989 Left GGTACTGGTGGAGTATTTGATAGTG 4990 Right TAACTTGAAACCCAAGGTACATTTC 4991 Left ATTACGATACACGTCTGCAGTCAAC 4992 Right ATGAAAATGGTCAGAGAAACCTTTA 4993 Left ACAGTTCATTACGATACACGTCTGC 4994 Right ATGAAAATGGTCAGAGAAACCTTTA 4995 Left TACAGTTCATTACGATACACGTCTG 4996 Right ATGAAAATGGTCAGAGAAACCTTTA 4997 Left TGGAGGAGTTTGTAAATGAAGTACAG 4998 Right TTTATCTGTATCAAAGAATGGTCCTG 4999 Left AGGTACTGGTGGAGTATTTGATAGTG 5000 Right TAACTTGAAACCCAAGGTACATTTC KRAS Exon2 401-600 bases 5001 Left AGTCATGATATGATCCTTTGAGAGC 5002 Right ATGAAAATGGTCAGAGAAACCTTTA 5003 Left ATATGATCCTTTGAGAGCCTTTAGC 5004 Right GAAACCCAAGGTACATTTCAGATAA 5005 Left TGATATGATCCTTTGAGAGCCTTTA 5006 Right ATGAAAATGGTCAGAGAAACCTTTA 5007 Left ATATGATCCTTTGAGAGCCTTTAGC 5008 Right ATGAAAATGGTCAGAGAAACCTTTA 5009 Left TGAAGTCATGATATGATCCTTTGAG 5010 Right ATGAAAATGGTCAGAGAAACCTTTA 5011 Left TGGAGGAGTTTGTAAATGAAGTACAG 5012 Right CCAAGGAAAGTAAAGTTCCCATATT 5013 Left TGGAGGAGTTTGTAAATGAAGTACAG 5014 Right TGACATACTCCCAAGGAAAGTAAAG 5015 Left TGGAGGAGTTTGTAAATGAAGTACAG 5016 Right CTGACATACTCCCAAGGAAAGTAAA 5017 Left ATGATATGATCCTTTGAGAGCCTTT 5018 Right ATGAAAATGGTCAGAGAAACCTTTA 5019 Left AGTCATGATATGATCCTTTGAGAGC 5020 Right TTTATCTGTATCAAAGAATGGTCCTG 5021 Left AGTCATGATATGATCCTTTGAGAGC 5022 Right TACTCATGAAAATGGTCAGAGAAAC 5023 Left TGGAGGAGTTTGTAAATGAAGTACAG 5024 Right TAACTTGAAACCCAAGGTACATTTC 5025 Left TGGAGGAGTTTGTAAATGAAGTACAG 5026 Right GAAACCCAAGGTACATTTCAGATAA 5027 Left AGTCATGATATGATCCTTTGAGAGC 5028 Right TTATCTGTATCAAAGAATGGTCCTG 5029 Left ATATGATCCTTTGAGAGCCTTTAGC 5030 Right TGAAACCCAAGGTACATTTCAGATA 5031 Left GATATGATCCTTTGAGAGCCTTTAG 5032 Right GAAACCCAAGGTACATTTCAGATAA 5033 Left TGAAGTACAGTTCATTACGATACACG 5034 Right CCAAGGAAAGTAAAGTTCCCATATT 5035 Left TGAAGTACAGTTCATTACGATACACG 5036 Right CGAAACTCTGAAATACACTTCCAAT 5037 Left TGAAGTACAGTTCATTACGATACACG 5038 Right CTGACATACTCCCAAGGAAAGTAAA 5039 Left TGAAGTACAGTTCATTACGATACACG 5040 Right TGACATACTCCCAAGGAAAGTAAAG KRAS Exon2 601-800 bases 5041 Left AGTCATGATATGATCCTTTGAGAGC 5042 Right CCAAGGAAAGTAAAGTTCCCATATT 5043 Left AGTCATGATATGATCCTTTGAGAGC 5044 Right CGAAACTCTGAAATACACTTCCAAT 5045 Left AGTCATGATATGATCCTTTGAGAGC 5046 Right TGACATACTCCCAAGGAAAGTAAAG 5047 Left AGTCATGATATGATCCTTTGAGAGC 5048 Right CTGACATACTCCCAAGGAAAGTAAA 5049 Left AGTCATGATATGATCCTTTGAGAGC 5050 Right AATTTCTACCCTCTCACGAAACTCT 5051 Left AGTCATGATATGATCCTTTGAGAGC 5052 Right GATACAAATTTCTACCCTCTCACGA 5053 Left TCTGTAGCTGTTGCATATTGACTTC 5054 Right CCAAGGAAAGTAAAGTTCCCATATT 5055 Left TTTTTCTGTAGCTGTTGCATATTGA 5056 Right CCAAGGAAAGTAAAGTTCCCATATT 5057 Left AGTCATGATATGATCCTTTGAGAGC 5058 Right TAACTTGAAACCCAAGGTACATTTC 5059 Left AGTCATGATATGATCCTTTGAGAGC 5060 Right GAAACCCAAGGTACATTTCAGATAA 5061 Left CTACTGCCATGATGCTTTAAAAGTT 5062 Right TAACTTGAAACCCAAGGTACATTTC 5063 Left AGAGCACTGTGAAGTCTCTACATGA 5064 Right CCAAGGAAAGTAAAGTTCCCATATT 5065 Left AGTCATGATATGATCCTTTGAGAGC 5066 Right TACAAATTTCTACCCTCTCACGAAA 5067 Left AGTCATGATATGATCCTTTGAGAGC 5068 Right AAATTTCTACCCTCTCACGAAACTC 5069 Left CTACTGCCATGATGCTTTAAAAGTT 5070 Right GAAACCCAAGGTACATTTCAGATAA 5071 Left AGAGCACTGTGAAGTCTCTACATGA 5072 Right CTGACATACTCCCAAGGAAAGTAAA 5073 Left AGAGCACTGTGAAGTCTCTACATGA 5074 Right TGACATACTCCCAAGGAAAGTAAAG 5075 Left TCTGTAGCTGTTGCATATTGACTTC 5076 Right TAACTTGAAACCCAAGGTACATTTC 5077 Left AGAGCACTGTGAAGTCTCTACATGA 5078 Right AATTTCTACCCTCTCACGAAACTCT 5079 Left AGTCATGATATGATCCTTTGAGAGC 5080 Right ACGAAACTCTGAAATACACTTCCAA KRAS Exon2 801-1000 bases 5081 Left ATCCAGCTTTATTTGACACTCATTC 5082 Right CCAAGGAAAGTAAAGTTCCCATATT 5083 Left CTACTGCCATGATGCTTTAAAAGTT 5084 Right CCAAGGAAAGTAAAGTTCCCATATT 5085 Left ATCCAGCTTTATTTGACACTCATTC 5086 Right CGAAACTCTGAAATACACTTCCAAT 5087 Left AATATTGTTCTTCTTTGCCTCAGTG 5088 Right TGACATACTCCCAAGGAAAGTAAAG 5089 Left AATATTGTTCTTCTTTGCCTCAGTG 5090 Right CTGACATACTCCCAAGGAAAGTAAA 5091 Left ATCCAGCTTTATTTGACACTCATTC 5092 Right TGACATACTCCCAAGGAAAGTAAAG 5093 Left ATCCAGCTTTATTTGACACTCATTC 5094 Right CTGACATACTCCCAAGGAAAGTAAA 5095 Left CTACTGCCATGATGCTTTAAAAGTT 5096 Right CGAAACTCTGAAATACACTTCCAAT 5097 Left CTACTGCCATGATGCTTTAAAAGTT 5098 Right TGACATACTCCCAAGGAAAGTAAAG 5099 Left CTACTGCCATGATGCTTTAAAAGTT 5100 Right CTGACATACTCCCAAGGAAAGTAAA 5101 Left ATCCAGCTTTATTTGACACTCATTC 5102 Right AATTTCTACCCTCTCACGAAACTCT 5103 Left ATCCAGCTTTATTTGACACTCATTC 5104 Right GATACAAATTTCTACCCTCTCACGA 5105 Left CTACTGCCATGATGCTTTAAAAGTT 5106 Right GATACAAATTTCTACCCTCTCACGA 5107 Left TCTGTAGCTGTTGCATATTGACTTC 5108 Right CGAAACTCTGAAATACACTTCCAAT 5109 Left TCTGTAGCTGTTGCATATTGACTTC 5110 Right TGACATACTCCCAAGGAAAGTAAAG 5111 Left TCTGTAGCTGTTGCATATTGACTTC 5112 Right AATTTCTACCCTCTCACGAAACTCT 5113 Left TCTGTAGCTGTTGCATATTGACTTC 5114 Right GATACAAATTTCTACCCTCTCACGA 5115 Left AAAAGTTTTTCTGTAGCTGTTGCAT 5116 Right CCAAGGAAAGTAAAGTTCCCATATT 5117 Left TTTTTCTGTAGCTGTTGCATATTGA 5118 Right CGAAACTCTGAAATACACTTCCAAT 5119 Left TTTTTCTGTAGCTGTTGCATATTGA 5120 Right TGACATACTCCCAAGGAAAGTAAAG KRAS Exon2 2 kb 5121 Left AGTCATGATATGATCCTTTGAGAGC 5122 Right TCCTAACCCACTTTATCACATTCAT 5123 Left AGTCATGATATGATCCTTTGAGAGC 5124 Right AATTAGACTGTTCCCCTTTACTGCT 5125 Left ATTATGTGTTACATGAGGGAAAGGA 5126 Right CCAAGGAAAGTAAAGTTCCCATATT 5127 Left AGTCATGATATGATCCTTTGAGAGC 5128 Right TTGGAAACAAAGTGTAATGGAATTT 5129 Left AGTCATGATATGATCCTTTGAGAGC 5130 Right TCAATTAGACTGTTCCCCTTTACTG 5131 Left ATTATGTGTTACATGAGGGAAAGGA 5132 Right CGAAACTCTGAAATACACTTCCAAT 5133 Left ATTATGTGTTACATGAGGGAAAGGA 5134 Right TGACATACTCCCAAGGAAAGTAAAG 5135 Left ATTATGTGTTACATGAGGGAAAGGA 5136 Right CTGACATACTCCCAAGGAAAGTAAA 5137 Left AATATTGTTCTTCTTTGCCTCAGTG 5138 Right TCTTTGCAAATAGGCATTATTTCTC 5139 Left ATCCAGCTTTATTTGACACTCATTC 5140 Right TCTTTGCAAATAGGCATTATTTCTC 5141 Left TCTTCAGTCAATTATGATGCTGTGT 5142 Right CCAAGGAAAGTAAAGTTCCCATATT 5143 Left GTCTTCAGTCAATTATGATGCTGTG 5144 Right CCAAGGAAAGTAAAGTTCCCATATT 5145 Left AATATTGTTCTTCTTTGCCTCAGTG 5146 Right AATTAGACTGTTCCCCTTTACTGCT 5147 Left ATCCAGCTTTATTTGACACTCATTC 5148 Right AATTAGACTGTTCCCCTTTACTGCT 5149 Left CTACTGCCATGATGCTTTAAAAGTT 5150 Right TCTTTGCAAATAGGCATTATTTCTC 5151 Left ATTATGTGTTACATGAGGGAAAGGA 5152 Right GATACAAATTTCTACCCTCTCACGA 5153 Left GCACATTCATTAATTTGGAGCTACT 5154 Right CTGACATACTCCCAAGGAAAGTAAA 5155 Left CTACTGCCATGATGCTTTAAAAGTT 5156 Right AATTAGACTGTTCCCCTTTACTGCT 5157 Left AATATTGTTCTTCTTTGCCTCAGTG 5158 Right ACCAAAAATATGTGACGTTTCCTTA 5159 Left AATATTGTTCTTCTTTGCCTCAGTG 5160 Right TACCAAAAATATGTGACGTTTCCTT KRAS Exon2 5 kb 5161 Left CAGCCAATAAGTCTAGGTAGAGCAG 5162 Right TAAAGATGAAACAAACCAATCCAAT 5163 Left AGCCTTCTTAAATCTTGAGATCCAT 5164 Right TCTTTGCAAATAGGCATTATTTCTC 5165 Left ATTATGTGTTACATGAGGGAAAGGA 5166 Right TAAAGATGAAACAAACCAATCCAAT 5167 Left GGCTAGTAAACTTTTTGGCCTTAAC 5168 Right TCTTTGCAAATAGGCATTATTTCTC 5169 Left AGCCTTCTTAAATCTTGAGATCCAT 5170 Right AATTAGACTGTTCCCCTTTACTGCT 5171 Left GGCTAGTAAACTTTTTGGCCTTAAC 5172 Right TCCTAACCCACTTTATCACATTCAT 5173 Left GGCTAGTAAACTTTTTGGCCTTAAC 5174 Right AATTAGACTGTTCCCCTTTACTGCT 5175 Left TTTGCTTTTAAGAGATGGTAGATGG 5176 Right TCTTTGCAAATAGGCATTATTTCTC 5177 Left CAGCCAATAAGTCTAGGTAGAGCAG 5178 Right TTGAACTGAATTATAAGTGCCACAA 5179 Left GGCTAGTAAACTTTTTGGCCTTAAC 5180 Right TTGGAAACAAAGTGTAATGGAATTT 5181 Left TCCTCATCTATTCCTAATTGCTTTG 5182 Right TTGAACTGAATTATAAGTGCCACAA 5183 Left TTTGCTTTTAAGAGATGGTAGATGG 5184 Right TCCTAACCCACTTTATCACATTCAT 5185 Left GCACATTCATTAATTTGGAGCTACT 5186 Right TAAAGATGAAACAAACCAATCCAAT 5187 Left GACTTAAACATGTGCATCTCCTTTT 5188 Right AAATGACAACAAAGCAAAGGTAAAG 5189 Left TCCTCATCTATTCCTAATTGCTTTG 5190 Right CCTTACTGAATAGGAAACTGTTCCA 5191 Left TTTGCTTTTAAGAGATGGTAGATGG 5192 Right AATTAGACTGTTCCCCTTTACTGCT 5193 Left AGTCATGATATGATCCTTTGAGAGC 5194 Right AATTAGCATGATTGCCTAGAAACAC 5195 Left AGCCTTCTTAAATCTTGAGATCCAT 5196 Right ACCAAAAATATGTGACGTTTCCTTA 5197 Left AGCCTTCTTAAATCTTGAGATCCAT 5198 Right TACCAAAAATATGTGACGTTTCCTT 5199 Left AGCCTTCTTAAATCTTGAGATCCAT 5200 Right TCAATTAGACTGTTCCCCTTTACTG

TABLE 11 ALK cDNA Capture Primer List for NGS Panel Seq. ID Primer Sequence ALK Region1 75-125 bases 5201 Left ctgcacactggccgtct 5202 Right aaccatgcttccctggagtg 5203 Left actgcacactggccg 5204 Right accatgcttccctgga 5205 Left actgcacactgg 5206 Right accatgcttccct ALK Region1 126-175 bases 5207 Left acaccatcctgagtccgtg 5208 Right cactgtccaaccatgcttcc 5209 Left gactccaagcacaccatcct 5210 Right aaccatgcttccctggagtg 5211 Left cctgagtccgtggatgagg 5212 Right ctggagcactgtccaaccat 5213 Left tccaagcacaccatcctgag 5214 Right ccaaccatgcttccctgga 5215 Left ctgcacactggccgtct 5216 Right ctcgaaatgggttgtctggac 5217 Left ccgtggatgaggagcagc 5218 Right gattcttccctggagcactgtc 5219 Left gagcactgcacactggc 5220 Right cgattcttccctggagcact 5221 Left atgaggagcagcagtgag 5222 Right gattcttccctggagcactgtccaa 5223 Left catcctgagtccgtggat 5224 Right agcactgtccaaccatgct 5225 Left tgagcactgcacact 5226 Right cccgattcttccctggagc 5227 Left atgaggagcagcagt 5228 Right gacgcccgattcttccct 5229 Left gtgagcactgcac 5230 Right aatgggttgtctggacgcc ALK Region1 176-225 bases 5231 Left ctcctttctccttctcaacacct 5232 Right attcttccctggagcactgtc 5233 Left cttctcaacacctcagctgact 5234 Right cgattcttccctggagcact 5235 Left tttctccttctcaacacctcagct 5236 Right gcactgtccaaccatgcttc 5237 Left ctgactccaagcacaccatc 5238 Right ctcgaaatgggttgtctggac 5239 Left tccaagcacaccatcctgag 5240 Right ccactcgaaatgggttgtctg 5241 Left cctgagtccgtggatgagg 5242 Right gagatgtattccagggccactc 5243 Left ttctcaacacctcagctgactccaa 5244 Right gattcttccctggagcactgtccaa 5245 Left gaggctcctttctccttctca 5246 Right ctggagcactgtccaacca 5247 Left ctcagctgactccaagcaca 5248 Right gagcactgtccaaccatgc 5249 Left acaccatcctgagtccgtg 5250 Right ttccagggccactcgaaat 5251 Left accatcctgagtccgtggat 5252 Right ggccactcgaaatgggttg 5253 Left ccgtggatgaggagcagc 5254 Right ggagatgtattccagggcca 5255 Left gagcactgcacactggc 5256 Right gacaagctgcggtttccac 5257 Left gcacaccatcctgagtcc 5258 Right gaaatgggttgtctggacgcc 5259 Left atgaggagcagcagtgag 5260 Right agggccactcgaaatggg 5261 Left ccgtggatgaggagc 5262 Right cccgattcttccctggagc 5263 Left tgagcactgcacact 5264 Right aagctgcggtttccactgg 5265 Left atgaggagcagcagt 5266 Right ggacgcccgattcttccc 5267 Left gtgagcactgcac 5268 Right tccactggagatgtattcca 5269 Left ttctccttctcaaca 5270 Right gcccgattcttccctgg ALK Region1 226-275 bases 5271 Left ctcctttctccttctcaacacct 5272 Right ctcgaaatgggttgtctggac 5273 Left gaggctcctttctccttctcaa 5274 Right ccactcgaaatgggttgtctg 5275 Left cttctcaacacctcagctgactc 5276 Right gagatgtattccagggccactc 5277 Left acaccatcctgagtccgtg 5278 Right caaagaagtccactgcagacaag 5279 Left ctccttctcaacacctcagct 5280 Right ggagatgtattccagggcca 5281 Left ccagaggctcctttctccttc 5282 Right ttccagggccactcgaaat 5283 Left cctgagtccgtggatgagg 5284 Right gcaaagaagtccactgcagac 5285 Left ttctcaacacctcagctgactccaa 5286 Right ggccactcgaaatgggttg 5287 Left gagcactgcacactggc 5288 Right gttccttcactgcagttcttcag 5289 Left ctgactccaagcacaccatc 5290 Right caagctgcggtttccactg 5291 Left tccaagcacaccatcctgag 5292 Right agacaagctgcggtttcca 5293 Left ctcagctgactccaagcaca 5294 Right gaaatgggttgtctggacgcc 5295 Left ccgtggatgaggagcagc 5296 Right ctgcagttcttcagggcaaag 5297 Left accatcctgagtccgtggat 5298 Right ggcaaagaagtccactgca 5299 Left atgaggagcagcagtgag 5300 Right cttcactgcagttcttcagggc 5301 Left cccagaggctcctttctcc 5302 Right gggccactcgaaatggg 5303 Left gcacaccatcctgagtcc 5304 Right tccactggagatgtattcca 5305 Left gtgagcactgcaca 5306 Right gatgttccttcactgcagttctt 5307 Left ccgtggatgaggagc 5308 Right ttcagggcaaagaagtcc 5309 Left tgaggagcagcagt 5310 Right tgttccttcactgcagtt ALK Region2 75-125 bases 5311 Left cactccagggaagcatggt 5312 Right caaagaagtccactgcagacaag 5313 Left aacgaggctgcaagagagat 5314 Right gagatgtattccagggccact 5315 Left gaggctgcaagagagatcct 5316 Right tggagatgtattccagggcc 5317 Left ctcctgatgcccactccag 5318 Right caagctgcggtttccactg 5319 Left tcctcctgatgcccactc 5320 Right agacaagctgcggtttcca 5321 Left acgaggctgcaagaga 5322 Right tccactggagatgtattcca 5323 Left cactccagggaagcat 5324 Right caaagaagtccactgcagac 5325 Left tcctcctgatgccca 5326 Right agacaagctgcggttt 5327 Left ccactccagggaag 5328 Right aaagaagtccactgca 5329 Left acgaggctgcaag 5330 Right tccactggagatgtatt 5331 Left agagatcctcctga 5332 Right tccactggagatgt ALK Region2 126-175 bases 5333 Left aacgaggctgcaagagagat 5334 Right caaagaagtccactgcagacaag 5335 Left ctcctgatgcccactccag 5336 Right gatgttccttcactgcagttctt 5337 Left gaggctgcaagagagatcct 5338 Right cttcactgcagttcttcaggg 5339 Left cacaacgaggctgcaagag 5340 Right gcaaagaagtccactgcagac 5341 Left ctctggaaggtacattgcccag 5342 Right caagctgcggtttccactg 5343 Left gaaggtacattgcccagctg 5344 Right agacaagctgcggtttcca 5345 Left tcctcctgatgcccactc 5346 Right ctgcagttcttcagggcaaag 5347 Left cccacaacgaggctgcaa 5348 Right ggcaaagaagtccactgca 5349 Left tgctgccccacaacgag 5350 Right ttcagggcaaagaagtcc 5351 Left gtacattgcccagctgctg 5352 Right agacaagctgcggttt 5353 Left tcctcctgatgccca 5354 Right gatgttccttcactgcagtt 5355 Left gccccacaacgaggct 5356 Right tccactggagatgtattc 5357 Left gctgctgccccacaac 5358 Right ggcaaagaagtccact 5359 Left ctctggaaggtacattgcc 5360 Right agacaagctgcgg 5361 Left cacctgcagccctct 5362 Right tccactggagatgta 5363 Left agagatcctcctga 5364 Right gatgttccttcactgca 5365 Left ctgctgccccac 5366 Right tcagggcaaagaag ALK Region2 176-225 bases 5367 Left gaaggtacattgcccagctg 5368 Right gatgttccttcactgcagttat 5369 Left gaggctgcaagagagatcct 5370 Right gttccttcactgcagttcttcag 5371 Left gtacattgcccagctgctg 5372 Right caaagaagtccactgcagacaag 5373 Left ctcctgatgcccactccag 5374 Right cattccaacaagtgaaggagctc 5375 Left ctctggaaggtacattgccca 5376 Right actgcagttcttcagggcaa 5377 Left cacaacgaggctgcaagagagat 5378 Right ctgcagttcttcagggcaaaga 5379 Left tcctcctgatgcccactc 5380 Right cccattccaacaagtgaaggag 5381 Left acaacgaggctgcaagaga 5382 Right atcttggagcctggggatgttc 5383 Left ccacaacgaggctgcaag 5384 Right atcttggagcctggggatg 5385 Left gagcactgcacactggc 5386 Right gacaagctgcggtttccac 5387 Left actgcacactggccgtc 5388 Right aagctgcggtttccactgg 5389 Left tgctgccccacaacgag 5390 Right gctctgcagggccatct 5391 Left gccccacaacgaggct 5392 Right gatgttccttcactgcagtt 5393 Left cacctgcagccctct 5394 Right gcaaagaagtccactgcagac 5395 Left tcctcctgatgccca 5396 Right gactgtcccattccaacaagtg 5397 Left ctctggaaggtacattgc 5398 Right ggcaaagaagtccactgca 5399 Left gctgctgccccacaac 5400 Right ttcagggcaaagaagtcc 5401 Left cgtctcggtgcacagg 5402 Right agacaagctgcggtttc 5403 Left ccgtggatgaggagcagc 5404 Right tccactggagatgtattc 5405 Left agagatcctcctga 5406 Right caacaagtgaaggagctctgc ALK Region2 226-275 bases 5407 Left ctctggaaggtacattgcccag 5408 Right cattccaacaagtgaaggagctc 5409 Left aacgaggctgcaagagagatc 5410 Right cccattccaacaagtgaaggag 5411 Left acaccatcctgagtccgtg 5412 Right caaagaagtccactgcagacaag 5413 Left gaaggtacattgcccagctg 5414 Right caacaagtgaaggagctctgc 5415 Left gtacattgcccagctgctg 5416 Right gactgtcccattccaacaagtg 5417 Left ctgcacactggccgtct 5418 Right gatgttccttcactgcagttctt 5419 Left cctgagtccgtggatgagg 5420 Right gcaaagaagtccactgcagac 5421 Left gagcactgcacactggc 5422 Right gttccttcactgcagttcttcag 5423 Left ctgactccaagcacaccatc 5424 Right caagctgcggtttccactg 5425 Left tccaagcacaccatcctgag 5426 Right agacaagctgcggtttcca 5427 Left ccgtggatgaggagcagc 5428 Right ctgcagttcttcagggcaaag 5429 Left accatcctgagtccgtggat 5430 Right ggcaaagaagtccactgca 5431 Left tcctgatgcccactccag 5432 Right ctcatcttctccctgggca 5433 Left cacaacgaggctgcaagag 5434 Right ggaagtcacaggcctgcc 5435 Left atgaggagcagcagtgag 5436 Right cttcactgcagttcttcagggc 5437 Left ctctggaaggtacattgcc 5438 Right atcttggagcctggggatgttc 5439 Left ctgctgccccacaacga 5440 Right tgaaggagctctgcaggg 5441 Left cgtctcggtgcacagg 5442 Right atcttggagcctggggatg 5443 Left gccccacaacgaggct 5444 Right gaaggagctctgcagggccatc 5445 Left cccacaacgaggctgcaa 5446 Right gactgtcccattccaacaa ALK Region3 76-125 bases 5447 Left ggaatacatctccagtggaaacc 5448 Right cattccaacaagtgaaggagctc 5449 Left gtccagacaacccatttcgag 5450 Right atcttggagcctggggatg 5451 Left tggccctggaatacatctcc 5452 Right acaagtgaaggagctctgca 5453 Left cgagtggccctggaatacatc 5454 Right tgaaggagctctgcaggg 5455 Left tcgagtggccctggaatac 5456 Right gctctgcagggccatct 5457 Left catttcgagtggccctgga 5458 Right gagctctgcagggcca 5459 Left gcgtccagacaacccatttc 5460 Right atcttggagcctgggg 5461 Left tggaatacatctccagtggaa 5462 Right attccaacaagtgaaggag 5463 Left ggcgtccagacaacccat 5464 Right catcttggagcctg 5465 Left ggaatacatctccagtg 5466 Right cattccaacaagtgaag ALK Region3 126-175 bases 5467 Left gtccagacaacccatttcgag 5468 Right cattccaacaagtgaaggagctc 5469 Left gagtggccctggaatacatctc 5470 Right cccattccaacaagtgaaggag 5471 Left cgagtggccctggaatacat 5472 Right gactgtcccattccaacaagtg 5473 Left gacagtgctccagggaagaat 5474 Right caacaagtgaaggagctctgc 5475 Left ggaagcatggttggacagtg 5476 Right gctctgcagggccatct 5477 Left tgctccagggaagaatcgg 5478 Right tgaaggagctctgcaggg 5479 Left gcgtccagacaacccatttc 5480 Right gagctctgcagggcca 5481 Left gaagaatcgggcgtccaga 5482 Right gactgtcccattccaacaa 5483 Left tcgagtggccctggaata 5484 Right ggaagtcacaggcctgcc 5485 Left catttcgagtggccctgga 5486 Right cccattccaacaagtgaag 5487 Left ggcgtccagacaacccat 5488 Right caagctggaggactgtc 5489 Left gaagaatcgggcgtccagacaa 5490 Right gactgtcccattccaa 5491 Left ccatttcgagtggccct 5492 Right caagctggaggact 5493 Left ggaagaatcgggcgtcc 5494 Right ggactgtcccattc 5495 Left ccatttcgagtggc 5496 Right ggcctgcccaag ALK Region3 176-225 bases 5497 Left gagtggccctggaatacatctc 5498 Right acatctggctctcatcttctcc 5499 Left gacagtgctccagggaagaat 5500 Right cattccaacaagtgaaggagctc 5501 Left ggaagcatggttggacagtg 5502 Right cccattccaacaagtgaaggag 5503 Left ccagacaacccatttcgagtg 5504 Right atctggctctcatcttctccctg 5505 Left cactccagggaagcatggtt 5506 Right gactgtcccattccaacaagtg 5507 Left gaggctgcaagagagatcct 5508 Right caacaagtgaaggagctctgc 5509 Left cgagtggccctggaataca 5510 Right gcacatctggctctcatcttc 5511 Left cgtccagacaacccatttcg 5512 Right ctctcatcttctccctgggc 5513 Left aacgaggctgcaagagagat 5514 Right gctctgcagggccatct 5515 Left gcatggttggacagtgctc 5516 Right tgaaggagctctgcaggg 5517 Left atggttggacagtgctccag 5518 Right ggaagtcacaggcctgcc 5519 Left agggaagcatggttggaca 5520 Right gagctctgcagggcca 5521 Left tgctccagggaagaatcgg 5522 Right gactgtcccattccaacaa 5523 Left tcgagtggccctggaat 5524 Right gcagtttccggcacatctg 5525 Left ctcctgatgcccactccag 5526 Right cccattccaacaagtgaag 5527 Left ccatttcgagtggccctgg 5528 Right gcacatctggctctcatc 5529 Left gaagaatcgggcgtccaga 5530 Right caagctggaggactgtc 5531 Left gaagaatcgggcgtccagacaac 5532 Right ttctccctgggcaca 5533 Left ggacagtgctccagggaag 5534 Right tggaagtcacaggcc 5535 Left agggaagaatcgggcgtc 5536 Right gactgtcccattccaa ALK Region3 226-275 bases 5537 Left gagtggccctggaatacatctc 5538 Right agccatcttcaaagttgcagtaaaa 5539 Left gtccagacaacccatttcgag 5540 Right aagccatcttcaaagttgcagta 5541 Left ctctggaaggtacattgcccag 5542 Right cattccaacaagtgaaggagctc 5543 Left gacagtgctccagggaagaatc 5544 Right acatctggctctcatcttctcc 5545 Left aacgaggctgcaagagagatc 5546 Right cccattccaacaagtgaaggag 5547 Left ggaagcatggttggacagtg 5548 Right gcacatctggctctcatcttc 5549 Left gaaggtacattgcccagctg 5550 Right caacaagtgaaggagctctgc 5551 Left gtacattgcccagctgctg 5552 Right gactgtcccattccaacaagtg 5553 Left gcgtccagacaacccatttc 5554 Right atctggctctcatcttctccctg 5555 Left cactccagggaagcatggtt 5556 Right ctctcatcttctccctgggc 5557 Left cgagtggccctggaatacat 5558 Right tccagccacagaagccatc 5559 Left atggttggacagtgctccag 5560 Right gcagtttccggcacatctg 5561 Left cacaacgaggctgcaagag 5562 Right ggaagtcacaggcctgcc 5563 Left catttcgagtggccctgga 5564 Right aagccatcttcaaagttgca 5565 Left gacagtgctccagggaaga 5566 Right cacaggcagtttccggc 5567 Left cccacaacgaggctgcaa 5568 Right tgaaggagctctgcaggg 5569 Left gcatggttggacagtgctc 5570 Right gcacatctggctctcatc 5571 Left tgctgccccacaacgag 5572 Right agctctgcagggccatc 5573 Left tcgagtggccctggaata 5574 Right gtccagccacagaagcc 5575 Left gccccacaacgaggct 5576 Right gactgtcccattccaacaa ALK Region4 226-275 bases 5577 Left gagcactgcacactggc 5578 Right ttggagcctggggatgttc 5579 Left ctgcacactggccgtct 5580 Right atcttggagcctggggatg 5581 Left cgtctcggtgcacagg 5582 Right gctctgcagggccatct 5583 Left cgtctcggtgcac 5584 Right agctctgcagggcca 5585 Left tgagcactgcacact 5586 Right tggagcctgggg 5587 Left aggagcagcagtgag 5588 Right gatgttccttcact ALK Region4 276-325 bases 5589 Left ctgcacactggccgtct 5590 Right cattccaacaagtgaaggagctc 5591 Left ctgactccaagcacaccatc 5592 Right atcttggagcctggggatgtt 5593 Left cactgcacactggccg 5594 Right cccattccaacaagtgaaggag 5595 Left atgaggagcagcagtgag 5596 Right caacaagtgaaggagctctgc 5597 Left cctgagtccgtggatgagg 5598 Right gctctgcagggccatct 5599 Left tccaagcacaccatcctgag 5600 Right atcttggagcctggggat 5601 Left ccatcctgagtccgtggat 5602 Right gagctctgcagggcca 5603 Left tgagcactgcacactgg 5604 Right ctgtcccattccaacaagtg 5605 Left cgtctcggtgcacagg 5606 Right tgaaggagctctgcaggg 5607 Left acaccatcctgagtccgtg 5608 Right atcttggagcctggg 5609 Left tgagcactgcacac 5610 Right gactgtcccattccaacaa 5611 Left cgtctcggtgcac 5612 Right cccattccaacaagtgaag 5613 Left ttctcaacacctcagctgactc 5614 Right gatgttccttcact 5615 Left ccgtggatgaggagcagc 5616 Right catcttggagcct 5617 Left gtgagcactgca 5618 Right gactgtcccattccaa ALK Region4 326-375 bases 5619 Left cttctcaacacctcagctgactc 5620 Right cattccaacaagtgaaggagctc 5621 Left aacacctcagctgactccaag 5622 Right cccattccaacaagtgaaggag 5623 Left tttctccttctcaacacctcagct 5624 Right caacaagtgaaggagctctgc 5625 Left ctcagctgactccaagcaca 5626 Right gactgtcccattccaacaagtg 5627 Left cctttctccttctcaacacctca 5628 Right tcttggagcctggggatg 5629 Left gctcctttctccttctcaacac 5630 Right agctctgcagggccatc 5631 Left tgcacactggccgtctc 5632 Right ctggctctcatcttctccctg 5633 Left acaccatcctgagtccgtg 5634 Right tgaaggagctctgcaggg 5635 Left cactgcacactggccgt 5636 Right ctctcatcttctccctgggc 5637 Left cctgagtccgtggatgagg 5638 Right ggaagtcacaggcctgcc 5639 Left gactccaagcacaccatcct 5640 Right gactgtcccattccaacaa 5641 Left gaggctcctttctccttctcaa 5642 Right atcttggagcctgggg 5643 Left accatcctgagtccgtggat 5644 Right cccattccaacaagtgaag 5645 Left tccaagcacaccatcctgag 5646 Right caagctggaggactgtc 5647 Left gagcactgcacactggc 5648 Right ttctccctgggcaca 5649 Left ccagaggctcctttctccttc 5650 Right gcagggccatcttg 5651 Left gctgactccaagcacaccat 5652 Right gactgtcccattccaa 5653 Left ccgtggatgaggagcagc 5654 Right tggaagtcacaggcc 5655 Left cccagaggctcctttctcc 5656 Right catcttggagcctg 5657 Left gcacaccatcctgagtcc 5658 Right caagctggaggact ALK Region4 376-425 bases 5659 Left ctcctttctccttctcaacacct 5660 Right gactgtcccattccaacaagtg 5661 Left gaggctcctttctccttctcaac 5662 Right cattccaacaagtgaaggagctc 5663 Left ctccttctcaacacctcagct 5664 Right cccattccaacaagtgaaggag 5665 Left ccagaggctcctttctccttc 5666 Right caacaagtgaaggagctctgc 5667 Left acaccatcctgagtccgtg 5668 Right acatctggctctcatcttctcc 5669 Left ctccaagcacaccatcctga 5670 Right ctggctctcatcttctccctg 5671 Left accatcctgagtccgtggat 5672 Right gcacatctggctctcatcttc 5673 Left ctgactccaagcacaccatc 5674 Right ctctcatcttctccctgggc 5675 Left cttctcaacacctcagctgactc 5676 Right ggaagtcacaggcctgcc 5677 Left cctgagtccgtggatgagg 5678 Right gcagtttccggcacatctg 5679 Left cccagaggctcctttctcc 5680 Right aaggagctctgcagggc 5681 Left ttctcaacacctcagctgactccaa 5682 Right gactgtcccattccaacaa 5683 Left ccgtggatgaggagcagc 5684 Right cacaggcagtttccggc 5685 Left ctcagctgactccaagcaca 5686 Right caagctggaggactgtc 5687 Left tggggcagagcgttct 5688 Right cccattccaacaagtgaag 5689 Left gcacaccatcctgagtcc 5690 Right ttctccctgggcaca 5691 Left ccgtggatgaggagc 5692 Right gcacatctggctctcatc 5693 Left agagcgttctaaggagatg 5694 Right gactgtcccattccaa 5695 Left atgaggagcagcagt 5696 Right gcagtttccggcacat 5697 Left cccagaggctcctttc 5698 Right caagctggaggact ALK Region4 426-475 bases 5699 Left ctcctttctccttctcaacacctc 5700 Right acatctggctctcatcttctcc 5701 Left gactccaagcacaccatcct 5702 Right agccatcttcaaagttgcagtaaaa 5703 Left gctcctttctccttctcaacac 5704 Right atctggctctcatcttctccctg 5705 Left cttctcaacacctcagctgactc 5706 Right gcacatctggctctcatcttc 5707 Left acaccatcctgagtccgtg 5708 Right aagccatcttcaaagttgcagta 5709 Left agatggacttgctggatggg 5710 Right cattccaacaagtgaaggagctc 5711 Left gaggctcctttctccttctcaa 5712 Right ctctcatcttctccctgggc 5713 Left tttctccttctcaacacctcagct 5714 Right gcagtttccggcacatctg 5715 Left ccgcatcccctccgag 5716 Right cccattccaacaagtgaaggag 5717 Left cctgagtccgtggatgagg 5718 Right tccagccacagaagccatc 5719 Left ccagaggctcctttctccttc 5720 Right ggaagtcacaggcctgcc 5721 Left ttctcaacacctcagctgactccaa 5722 Right cacaggcagtttccggc 5723 Left tccaagcacaccatcctgag 5724 Right aagccatcttcaaagttgca 5725 Left cagagctggtcctggcg 5726 Right aacaagtgaaggagctctgca 5727 Left cagatggacttgctggat 5728 Right gactgtcccattccaacaagtg 5729 Left ccatcctgagtccgtggat 5730 Right gtccagccacagaagcc 5731 Left ccgtggatgaggagcagc 5732 Right tgtgccttgggtccagc 5733 Left ctcagctgactccaagcaca 5734 Right gcacatctggctctcatc 5735 Left tcctggcgccgcatc 5736 Right tgaaggagctctgcaggg 5737 Left gctgactccaagcacaccat 5738 Right gcagtttccggcacat ALK Region4 476-525 bases 5739 Left ctcctttctccttctcaacacct 5740 Right agccatcttcaaagttgcagtaaaa 5741 Left gaggctcctttctccttctcaac 5742 Right aagccatcttcaaagttgcagta 5743 Left gagtattcccctccactgcat 5744 Right cattccaacaagtgaaggagctc 5745 Left tattcccctccactgcatgac 5746 Right cccattccaacaagtgaaggag 5747 Left agatggacttgctggatggg 5748 Right acatctggctctcatcttctcc 5749 Left cactgcatgacctcaggaac 5750 Right gactgtcccattccaacaagtg 5751 Left gcatgacctcaggaaccaga 5752 Right caacaagtgaaggagctctgc 5753 Left cttctcaacacctcagctgactc 5754 Right tccagccacagaagccatc 5755 Left ctgactccaagcacaccatc 5756 Right attgaggagtgtggggtgac 5757 Left ctccttctcaacacctcagct 5758 Right tgacagtgtgccttgggtc 5759 Left ccagaggctcctttctccttc 5760 Right gcagtttccggcacatctg 5761 Left aacacctcagctgactccaag 5762 Right agtgtgccttgggtccag 5763 Left ctccaagcacaccatcctga 5764 Right agtgtggggtgacagtgtg 5765 Left gtattcccctccactgcatgacctc 5766 Right tgaaggagctctgcaggg 5767 Left acaccatcctgagtccgtg 5768 Right gtcctgacctgccattgag 5769 Left ctcagctgactccaagcaca 5770 Right ggggtgacagtgtgcctt 5771 Left cccagaggctcctttctcc 5772 Right aagccatcttcaaagttgca 5773 Left agagcgttctaaggagatg 5774 Right gcacatctggctctcatcttc 5775 Left accatcctgagtccgtggat 5776 Right gtccagccacagaagcc 5777 Left tccactgcatgacctcagg 5778 Right gactgtcccattccaacaa ALK Region4 750-1250 bases 5779 Left tggaatctcacctggataatgaaag 5780 Right ttttgttctccactagcaccaag 5781 Left gaatcaccaacaaacatgccttc 5782 Right agccatcttcaaagttgcagtaaaa 5783 Left cacctggataatgaaagactccttc 5784 Right tggtcactgtagcactttcagaa 5785 Left tggataatgaaagactccttccctt 5786 Right caatagagcatggtcttggtgg 5787 Left attttacatggaatctcacctggat 5788 Right gaaacgtagcactggtcactgtag 5789 Left atctcacctggataatgaaagactc 5790 Right cacccggttttgttctccactag 5791 Left tccttctcctgattattttacatgga 5792 Right aagccatcttcaaagttgcagta 5793 Left acatggaatctcacctggataatga 5794 Right gaaacgtagcactggtcactg 5795 Left gactggtcatagctccttggaatc 5796 Right ggttttgttctccactagcacc 5797 Left cagatcttcgggactggtcatag 5798 Right acatctggctctcatcttctcc 5799 Left cctgattattttacatggaatctcacc 5800 Right tgtcagacacatcgaggagag 5801 Left ctcctttctccttctcaacacctc 5802 Right ctccttcccggttttgttctc 5803 Left gctcctttctccttctcaacac 5804 Right tttgttctccactagcaccaaggac 5805 Left aaagactccttccctttcctgt 5806 Right ctcaagactccacgaatgagc 5807 Left aatcaccaacaaacatgccttctcc 5808 Right ctagcaccaaggacacgtttc 5809 Left cttctccactcctgattattttaca 5810 Right cttcccggttttgttctccac 5811 Left tctcctgattattttacatggaatctc 5812 Right gtagcactggtcactgtagcacttt 5813 Left gactggtcatagctccttgga 5814 Right gcacatctggctctcatcttc 5815 Left gagaagaaggcgtcggaagt 5816 Right cattccaacaagtgaaggagctc 5817 Left gatcttcgggactggtcatagctc 5818 Right atctggctctcatcttctccctg

TABLE 12 EGFR cDNA Capture Primer List for NGS Panel Seq. ID Primer Sequence EGFR Region1 75-125 bases 5819 Left CCCAGTGGAGAAGCTC 5820 Right GGGATCCAGAGTCCCTTATACA 5821 Left GAGAAGCTCCCAACCA 5822 Right GGGATCCAGAGTCCCTTAT 5823 Left CCCAGTGGAGAAG 5824 Right TGGGATCCAGAGTCCC 5825 Left GAGAAGCTCCCAA 5826 Right TGGGATCCAGAGT EGFR Region1 126-175 bases 5827 Left TTGTGGAGCCTCTTACACCC 5828 Right ACGGGAATTTTAACTTTCTCACCTT 5829 Left CTTGTGGAGCCTCTTACACCCAGT 5830 Right TGATAGCGACGGGAATTTTAACT 5831 Left GGAGAGGGAGCTTGTGGAG 5832 Right GGGATCCAGAGTCCCTTATACA 5833 Left CTTGTGGAGCCTCTTACA 5834 Right CGACGGGAATTTTAACTTTCTCAC 5835 Left CTGCAGGAGAGGGAGCTTG 5836 Right AATTTTAACTTTCTCACCTTCTG 5837 Left CCCAGTGGAGAAGCTC 5838 Right TTCTCTTAATTCCTTGATAGCGACG 5839 Left GAGAAGCTCCCAACCA 5840 Right CTTAATTCCTTGATAGCGACGGGAA 5841 Left TGCTGCAGGAGAGGGA 5842 Right GGGATCCAGAGTCCCTTAT 5843 Left CTTGTGGAGCCTCTT 5844 Right CCTTGATAGCGACGGGAATTTTA 5845 Left GAGAAGCTCCCAA 5846 Right GCGACGGGAATTTTAACTTTCT 5847 Left GCACGCTGCGGA 5848 Right TGGGATCCAGAGTCCC 5849 Left CCCAGTGGAGAAG 5850 Right TCTCTTAATTCCTTGATAGCG 5851 Left CGGAGGCTGCTG 5852 Right TGGGATCCAGAGT 5853 Left TGCTGCAGGAGAG 5854 Right CTTCTGGGATCCA EGFR Region1 176-225 bases 5855 Left TTGTGGAGCCTCTTACACCC 5856 Right TTCTCTTAATTCCTTGATAGCGACG 5857 Left CAGGAGAGGGAGCTTGTGG 5858 Right ACGGGAATTTTAACTTTCTCACCTT 5859 Left CTGCAGGAGAGGGAGCTTG 5860 Right TGATAGCGACGGGAATTTTAACTT 5861 Left CTTGTGGAGCCTCTTACACCCAGT 5862 Right CTTAATTCCTTGATAGCGACGGGAA 5863 Left CCACATCGTTCGGAAGCG 5864 Right CGACGGGAATTTTAACTTTCTCAC 5865 Left CATCGTTCGGAAGCGCAC 5866 Right CCTTGATAGCGACGGGAATTTTAA 5867 Left ATCGGCCTCTTCATGCGAA 5868 Right GGGATCCAGAGTCCCTTATACA 5869 Left GAAGGCGCCACATCGTTC 5870 Right GCGACGGGAATTTTAACTTTCT 5871 Left TGCTGCAGGAGAGGGAG 5872 Right TCCTTGATAGCGACGGGAATTT 5873 Left GCGCCACATCGTTCGGAA 5874 Right AATTTTAACTTTCTCACCTTCTG 5875 Left GGATCGGCCTCTTCATGC 5876 Right GGGATCCAGAGTCCCTTAT 5877 Left CCCAGTGGAGAAGCTC 5878 Right ACGTAGGCTTCATCGAGGATTT 5879 Left CTTGTGGAGCCTCTTACA 5880 Right CCTTGTTGGCTTTCGGAGA 5881 Left GAGAAGCTCCCAACCA 5882 Right CACGTAGGCTTCATCGAGGA 5883 Left CGAAGGCGCCACATCG 5884 Right TGGGATCCAGAGTCCC 5885 Left TGCTGCAGGAGAGG 5886 Right AGATGTTGCTTCTCTTAATTCC 5887 Left GCACGCTGCGGA 5888 Right TTCTCTTAATTCCTTGATAGCG 5889 Left CCCAGTGGAGAAG 5890 Right GGCCATCACGTAGGCTTCAT 5891 Left CTTGTGGAGCCTCTT 5892 Right TCGGAGATGTTGCTTCT 5893 Left GAGAAGCTCCCAA 5894 Right CTGGCCATCACGTAGGCTT EGFR Region1 226-275 bases 5895 Left ATCGGCCTCTTCATGCGAA 5896 Right TTCTCTTAATTCCTTGATAGCGACG 5897 Left CCTCCTCTTGCTGCTGGT 5898 Right ACGGGAATTTTAACTTTCTCACCTT 5899 Left TTGTGGAGCCTCTTACACCC 5900 Right CGTAGGCTTCATCGAGGATTTC 5901 Left GAAGGCGCCACATCGTTC 5902 Right TGATAGCGACGGGAATTTTAACTT 5903 Left CCTCTTGCTGCTGGTGGT 5904 Right CGACGGGAATTTTAACTTTCTCAC 5905 Left CCACATCGTTCGGAAGCG 5906 Right CTTAATTCCTTGATAGCGACGGGAA 5907 Left GGATCGGCCTCTTCATGC 5908 Right CCTTGATAGCGACGGGAATTTTAA 5909 Left GGAGAGGGAGCTTGTGGAG 5910 Right CACGTAGGCTTCATCGAGGAT 5911 Left GCCCTCCTCTTGCTGCT 5912 Right GGGATCCAGAGTCCCTTATACA 5913 Left CTTGTGGAGCCTCTTACACCCAGT 5914 Right GGCCATCACGTAGGCTTCAT 5915 Left CTGGTGGTGGCCCTGG 5916 Right TTCCTTGATAGCGACGGGAATTT 5917 Left CTGCAGGAGAGGGAGCTTG 5918 Right CTGGCCATCACGTAGGCTT 5919 Left CTGCTGGTGGTGGCCC 5920 Right GCGACGGGAATTTTAACTTTCT 5921 Left CATCGTTCGGAAGCGCAC 5922 Right CCTTGTTGGCTTTCGGAGA 5923 Left GCGCCACATCGTTCGGA 5924 Right AGATGTTGCTTCTCTTAATTCC 5925 Left TGCTGCAGGAGAGGGAG 5926 Right TCACGTAGGCTTCATCGAG 5927 Left CGAAGGCGCCACATCG 5928 Right TTCTCTTAATTCCTTGATAGCG 5929 Left GGGCCCTCCTCTTGCT 5930 Right GGGATCCAGAGTCCCTTAT 5931 Left CTTGTGGAGCCTCTTACA 5932 Right CGCTGGCCATCACGTAGG 5933 Left GAGAAGCTCCCAACCA 5934 Right GGTGGAGGTGAGGCAGATG EGFR Region2 75-125 bases 5935 Left AATCCTCGATGAAGCCTACGT 5936 Right CCAGGAGGCAGCCGAAG 5937 Left AATCCTCGATGAAGCCTACGTGATG 5938 Right AGGAGGCAGCCG 5939 Left GGAAATCCTCGATGAAGCCTA 5940 Right CGAAGGGCATGAG EGFR Region2 126-175 bases 5941 Left GAAATCCTCGATGAAGCCTACG 5942 Right GAGCCAATATTGTCTTTGTGTTCC 5943 Left GAAATCCTCGATGAAGCCTACGTGA 5944 Right AATATTGTCTTTGTGTTCCCGGAC 5945 Left AACATCTCCGAAAGCCAACAAG 5946 Right CTTTGTGTTCCCGGACATAGTC 5947 Left CTCGATGAAGCCTACGTGATGG 5948 Right ATTGTCTTTGTGTTCCCGGACATA 5949 Left TCGCTATCAAGGAATTAAGAGAAGC 5950 Right CCAGGAGGCAGCCGAAG 5951 Left GGAAATCCTCGATGAAGCC 5952 Right GGGAGCCAATATTGTCTTTGTGT 5953 Left CAAGGAATTAAGAGAAGCAACATCT 5954 Right GACATAGTCCAGGAGG 5955 Left CCGTCGCTATCAAGGAATTAAGAG 5956 Right AGGAGGCAGCCG 5957 Left TTAAAATTCCCGTCGCTATCAAGG 5958 Right CGAAGGGCATGAG 5959 Left GGAAATCCTCGATGAA 5960 Right TGGGAGCCAATATTGTCTTTG 5961 Left CAACAAGGAAATCC 5962 Right TGGGAGCCAATATTGTCT EGFR Region2 176-225 bases 5963 Left GAAAGTTAAAATTCCCGTCGCTATC 5964 Right GAGCCAATATTGTCTTTGTGTTCC 5965 Left GAAGGTGAGAAAGTTAAAATTCCCG 5966 Right ATTGTCTTTGTGTTCCCGGACATAG 5967 Left GTGAGAAAGTTAAAATTCCCGTCG 5968 Right AATATTGTCTTTGTGTTCCCGGAC 5969 Left CCGTCGCTATCAAGGAATTAAGAG 5970 Right GGGAGCCAATATTGTCTTTGTGT 5971 Left GAAATCCTCGATGAAGCCTACG 5972 Right GACGGTCCTCCAAGTAGTTCAT 5973 Left TCGCTATCAAGGAATTAAGAGAAGC 5974 Right TGGGAGCCAATATTGTCTTTG 5975 Left AGAAGCAACATCTCCGAAAGC 5976 Right GATCTGCACACACCAGTTGAG 5977 Left GAAATCCTCGATGAAGCCTACGTGA 5978 Right CGACGGTCCTCCAAGTAGTT 5979 Left CTCGATGAAGCCTACGTGATGG 5980 Right TCCTCCAAGTAGTTCATGCCC 5981 Left CAACATCTCCGAAAGCCAACAA 5982 Right CACACACCAGTTGAGCAGG 5983 Left TTAAAATTCCCGTCGCTATCAAGG 5984 Right CCAGGAGGCAGCCGAAG 5985 Left GAAGCAACATCTCCGAAAGCCAA 5986 Right CGATCTGCACACACCAGTT 5987 Left AATTCCCGTCGCTATCAAGGAATTA 5988 Right TGGGAGCCAATATTGTCT 5989 Left AAATCCTCGATGAAGCCT 5990 Right CCTCCAAGTAGTTCATGCCCTTT 5991 Left TCAAGGAATTAAGAGAAGCAACATCT 5992 Right GACATAGTCCAGGAGG 5993 Left TATCAAGGAATTAAGAGAAGCAACA 5994 Right GGTACTGGGAGCCA 5995 Left CCCAGAAGGTGAGAAAGTTAAAAT 5996 Right AGGAGGCAGCCG 5997 Left AGAAGCAACATCTCCGAA 5998 Right CGATCTGCACACACCA 5999 Left GAAATCCTCGATGAAG 6000 Right GCGACGGTCCTCCAAGTA 6001 Left GGCACGGTGTATAAGGGAC 6002 Right CGAAGGGCATGAG EGFR Region2 226-275 bases 6003 Left AAGGTGAGAAAGTTAAAATTCCCGT 6004 Right GGAGCCAATATTGTCTTTGTGTTC 6005 Left GAAAGTTAAAATTCCCGTCGCTATC 6006 Right TGGGAGCCAATATTGTCTTTGTG 6007 Left TCGCTATCAAGGAATTAAGAGAAGC 6008 Right GACGGTCCTCCAAGTAGTTCAT 6009 Left GTGAGAAAGTTAAAATTCCCGTCGC 6010 Right CCAATATTGTCTTTGTGTTCCCG 6011 Left CCGTCGCTATCAAGGAATTAAGAG 6012 Right CCTCCAAGTAGTTCATGCCCTTT 6013 Left TCAAGGAATTAAGAGAAGCAACATCT 6014 Right TCCTCCAAGTAGTTCATGCCC 6015 Left TTAAAATTCCCGTCGCTATCAAGG 6016 Right GATCTGCACACACCAGTTGAG 6017 Left GAAATCCTCGATGAAGCCTACG 6018 Right TGTTTTCACCAGTACGTTCCTG 6019 Left AGAAGGTGAGAAAGTTAAAATTCC 6020 Right ATATTGTCTTTGTGTTCCCGGAC 6021 Left TATCAAGGAATTAAGAGAAGCAACA 6022 Right CGACGGTCCTCCAAGTAGTT 6023 Left ATTCCCGTCGCTATCAAGGAATTAA 6024 Right CACACACCAGTTGAGCAGG 6025 Left CCCAGAAGGTGAGAAAGTTAAAAT 6026 Right GTCTTTGTGTTCCCGGACATA 6027 Left AATTCCCGTCGCTATCAAGGAAT 6028 Right CGATCTGCACACACCAGTT 6029 Left AACATCTCCGAAAGCCAACAAG 6030 Right AAGCGACGGTCCTCCAAGTA 6031 Left GGCACGGTGTATAAGGGAC 6032 Right CTTTGTGTTCCCGGACATAGTC 6033 Left TCGATGAAGCCTACGTGATGG 6034 Right GACATGCTGCGGTGTTTTCA 6035 Left GAAATCCTCGATGAAGCCTACGTGA 6036 Right AGTACGTTCCTGGCTGCC 6037 Left AGAAGCAACATCTCCGAAAGC 6038 Right CAAGCGACGGTCCTCCAA 6039 Left AAAAGATCAAAGTGCTGGGCTC 6040 Right CCAGGAGGCAGCCGAAG 6041 Left GAAGCAACATCTCCGAAAGCCAAC 6042 Right CTGCCAGGTCGCGGT EGFR Region3 75-125 bases 6043 Left CAAAGGGCATGAACTACTTGGAG 6044 Right CCAGCAGTTTGGCC 6045 Left AAAGGGCATGAACTACTTGGAGGAC 6046 Right CACCCAGCAGTTTG 6047 Left GCATGAACTACTTG 6048 Right TTCCGCACCCAG EGFR Region3 126-175 bases 6049 Left CAAAGGGCATGAACTACTTGGAG 6050 Right CCTTCTGCATGGTATTCTTTCTCTT 6051 Left AAAGGGCATGAACTACTTGGAGGAC 6052 Right CCTCCTTCTGCATGGTATTCTTTC 6053 Left CAACTGGTGTGTGCAGATCG 6054 Right CTGCATGGTATTCTTTCTCTTCCG 6055 Left CTCCCAGTACCTGCTCAACT 6056 Right CTTTCTCTTCCGCACCCAG 6057 Left GCTCAACTGGTGTGTGC 6058 Right CATGGTATTCTTTCTCTTCCGCAC 6059 Left AAAGGGCATGAACTACTTG 6060 Right GCCTCCTTCTGCATGGTATTCT 6061 Left GGCTCCCAGTACCTGCTC 6062 Right CCAGCAGTTTGGCC 6063 Left CAAAGGGCATGAACTAC 6064 Right CACTTTGCCTCCTTCTGC 6065 Left GCTCAACTGGTGTGTGCAGA 6066 Right CACCCAGCAGTTTG 6067 Left CAGATCGCAAAGGG 6068 Right GCCTCCTTCTGCATGGTAT 6069 Left CAAAGGGCATGAA 6070 Right GCCTCCTTCTGCATGG EGFR Region3 176-225 bases 6071 Left CAAAGGGCATGAACTACTTGGAG 6072 Right CTCTGGTGGGTATAGATTCTGTGTA 6073 Left CTATGTCCGGGAACACAAAGAC 6074 Right CCTTCTGCATGGTATTCTTTCTCTT 6075 Left GACTATGTCCGGGAACACAAA 6076 Right CCTCCTTCTGCATGGTATTCTTTC 6077 Left AAAGGGCATGAACTACTTGGAGGAC 6078 Right ACTCTGGTGGGTATAGATTCTGT 6079 Left CTCCCAGTACCTGCTCAACT 6080 Right CTGCATGGTATTCTTTCTCTTCCG 6081 Left CTCAACTGGTGTGTGCAGATC 6082 Right GCCTCCTTCTGCATGGTATTCT 6083 Left GGCTCCCAGTACCTGCTC 6084 Right CATGGTATTCTTTCTCTTCCGCAC 6085 Left GTCCGGGAACACAAAGACAATATT 6086 Right GCCTCCTTCTGCATGGTAT 6087 Left CTGCTCAACTGGTGTGTGC 6088 Right TTCCAATGCCATCCACTTGAT 6089 Left GTCCGGGAACACAAAGACAAT 6090 Right TTTCTCTTCCGCACCCAG 6091 Left CAAAGGGCATGAACTACTTG 6092 Right AGATTCTGTGTAAAATTGATTCCA 6093 Left TGGCTCCCAGTACCTG 6094 Right CACTTTGCCTCCTTCTGC 6095 Left GACTATGTCCGGGAACAC 6096 Right GCCTCCTTCTGCATGG 6097 Left CTTCGGCTGCCTCCTGG 6098 Right CCAGCAGTTTGGCC 6099 Left AAAGGGCATGAACTAC 6100 Right ACTCTGGTGGGTATAGATTC 6101 Left CTGCTCAACTGGTGT 6102 Right TTCCAATGCCATCCACTT 6103 Left CCCTTCGGCTGCCTCC 6104 Right CACCCAGCAGTTTG 6105 Left ACAATATTGGCTCCCA 6106 Right ATCCACTTGATAGGCAC 6107 Left CAGATCGCAAAGGG 6108 Right AGATTCTGTGTAAAATTGAT 6109 Left CAAAGGGCATGAAC 6110 Right CTCTGGTGGGTATAGA EGFR Region3 226-275 bases 6111 Left GTCCGGGAACACAAAGACAATATT 6112 Right ATTCCAATGCCATCCACTTGAT 6113 Left CTCCCAGTACCTGCTCAACT 6114 Right CTCTGGTGGGTATAGATTCTGTGTA 6115 Left GCTCATCACGCAGCTCATG 6116 Right CCTTCTGCATGGTATTCTTTCTCTT 6117 Left CAACTGGTGTGTGCAGATCG 6118 Right ACTCTGGTGGGTATAGATTCTGT 6119 Left CAGCTCATCACGCAGCTC 6120 Right CCTCCTTCTGCATGGTATTCTTTC 6121 Left CTCCACCGTGCAGCTCAT 6122 Right CTGCATGGTATTCTTTCTCTTCCG 6123 Left CGTGCAGCTCATCACGC 6124 Right CATGGTATTCTTTCTCTTCCGCAC 6125 Left CTATGTCCGGGAACACAAAGAC 6126 Right GATTCTGTGTAAAATTGATTCCA 6127 Left GCTCAACTGGTGTGTGCAG 6128 Right CCGTAGCTCCAGACATCACT 6129 Left CTATGTCCGGGAACACAAAGACAAT 6130 Right CACTTTGCCTCCTTCTGC 6131 Left CTTCGGCTGCCTCCTGG 6132 Right GCCTCCTTCTGCATGGTATTCT 6133 Left CTGCCTCACCTCCACCG 6134 Right CTTTCTCTTCCGCACCCAG 6135 Left GACTATGTCCGGGAACACAAA 6136 Right ATTCCAATGCCATCCACTT 6137 Left CTGCTCAACTGGTGTGTG 6138 Right AAACAGTCACCCCGTAGCTC 6139 Left GCTCCCAGTACCTGCTCA 6140 Right CCCGTAGCTCCAGACATC 6141 Left CCTCCACCGTGCAGCT 6142 Right GCCTCCTTCTGCATGGTAT 6143 Left TGGCTCCCAGTACCTGC 6144 Right ACTCTGGTGGGTATAGATTC 6145 Left CCCTTCGGCTGCCTCC 6146 Right GCCTCCTTCTGCATGG 6147 Left CAGATCGCAAAGGG 6148 Right AAAGGTCATCAACTCCCAAACAG 6149 Left CTGCTCAACTGGTGT 6150 Right CACCCCGTAGCTCCAGAC EGFR Region4 276-325 bases 6151 Left CTCCCAACCAAGCTCTCTT 6152 Right TTGTGTTCCCGGACATAGTC 6153 Left GAGAAGCTCCCAACCAAG 6154 Right CCAGGAGGCAGCCGAAG 6155 Left TCTCTTGAGGATCTTGA 6156 Right GCCAATATTGTCTTTGTGTTCCC 6157 Left CTCCCAACCAAGCTCT 6158 Right TTGTGTTCCCGGACATA 6159 Left TTGTGGAGCCTCTTACACCC 6160 Right CGAAGGGCATGAG 6161 Left CCCAGTGGAGAAGCTC 6162 Right AGGAGGCAGCCG 6163 Left GAGAAGCTCCCAACC 6164 Right GACATAGTCCAGGAGG EGFR Region4 326-375 bases 6165 Left CTTGTGGAGCCTCTTACACCCAG 6166 Right GAGCCAATATTGTCTTTGTGTTCC 6167 Left AGGAGAGGGAGCTTGTGGA 6168 Right ATTGTCTTTGTGTTCCCGGACATAG 6169 Left CTCCCAACCAAGCTCTCTT 6170 Right AATATTGTCTTTGTGTTCCCGGAC 6171 Left CTTGTGGAGCCTCTTACACC 6172 Right TGGGAGCCAATATTGTCTTTGT 6173 Left CTGCAGGAGAGGGAGCTTG 6174 Right CCAGGAGGCAGCCGAAG 6175 Left GAGAAGCTCCCAACCAAG 6176 Right CACACACCAGTTGAGCAGG 6177 Left CTCCCAACCAAGCTCT 6178 Right GATCTGCACACACCAGTTGAG 6179 Left TCTCTTGAGGATCTTGA 6180 Right CGATCTGCACACACCAGTT 6181 Left CTTGTGGAGCCTCTTAC 6182 Right TGGGAGCCAATATTGTCTT 6183 Left TGCTGCAGGAGAGGGAG 6184 Right GACATAGTCCAGGAGG 6185 Left CCACATCGTTCGGAAGCG 6186 Right CGAAGGGCATGAG 6187 Left CCCAGTGGAGAAGCT 6188 Right CCAGTTGAGCAGGTAC 6189 Left GAGAAGCTCCCAACC 6190 Right TGGGAGCCAATATTGT 6191 Left TCGGAAGCGCACG 6192 Right AGGAGGCAGCCG 6193 Left TGTGGAGCCTCT 6194 Right TACTGGGAGCCA EGFR Region4 376-425 bases 6195 Left AGGAGAGGGAGCTTGTGGA 6196 Right GAGCCAATATTGTCTTTGTGTTCC 6197 Left GAAGGCGCCACATCGTTC 6198 Right AATATTGTCTTTGTGTTCCCGGAC 6199 Left TTGTGGAGCCTCTTACACCC 6200 Right CTCCAAGTAGTTCATGCCCTTT 6201 Left CCACATCGTTCGGAAGCG 6202 Right GGGAGCCAATATTGTCTTTGTGT 6203 Left CTGCAGGAGAGGGAGCTTG 6204 Right TTGTCTTTGTGTTCCCGGACATAG 6205 Left TGGAGCCTCTTACACCCAGT 6206 Right TCCTCCAAGTAGTTCATGCCC 6207 Left CTCCCAACCAAGCTCTCTT 6208 Right GACGGTCCTCCAAGTAGTTCAT 6209 Left TGCTGCAGGAGAGGGAG 6210 Right GATCTGCACACACCAGTTGAG 6211 Left CATCGTTCGGAAGCGCAC 6212 Right TGGGAGCCAATATTGTCTTTG 6213 Left ATCGGCCTCTTCATGCGAA 6214 Right CCAGGAGGCAGCCGAAG 6215 Left GAGAAGCTCCCAACCAAG 6216 Right CGACGGTCCTCCAAGTAGTT 6217 Left CTTGTGGAGCCTCTTACA 6218 Right CACACACCAGTTGAGCAGG 6219 Left CCCAGTGGAGAAGCTC 6220 Right AAGCGACGGTCCTCCAAGTA 6221 Left CGCCACATCGTTCGGAA 6222 Right TGGGAGCCAATATTGTCT 6223 Left CTCCCAACCAAGCTCT 6224 Right CAAGCGACGGTCCTCCAA 6225 Left GAGAAGCTCCCAACC 6226 Right CGATCTGCACACACCAGTT 6227 Left TCTCTTGAGGATCTTGA 6228 Right GTACGTTCCTGGCTGCCA 6229 Left GGATCGGCCTCTTCATGC 6230 Right GACATAGTCCAGGAGG 6231 Left CTGGTGGTGGCCCTGG 6232 Right AGGAGGCAGCCG 6233 Left CCTCTTGCTGCTGGTGGT 6234 Right CGAAGGGCATGAG EGFR Region4 426-475 bases 6235 Left ATCGGCCTCTTCATGCGAA 6236 Right GAGCCAATATTGTCTTTGTGTTCC 6237 Left CCTCCTCTTGCTGCTGGT 6238 Right AATATTGTCTTTGTGTTCCCGGAC 6239 Left TTGTGGAGCCTCTTACACCC 6240 Right GACGGTCCTCCAAGTAGTTCAT 6241 Left GAAGGCGCCACATCGTTC 6242 Right GGGAGCCAATATTGTCTTTGTGT 6243 Left CTTGTGGAGCCTCTTACACCCAGT 6244 Right CCTCCAAGTAGTTCATGCCCTTT 6245 Left CCTCTTGCTGCTGGTGGT 6246 Right ATTGTCTTTGTGTTCCCGGACATAG 6247 Left GGAGAGGGAGCTTGTGGAG 6248 Right TCCTCCAAGTAGTTCATGCCC 6249 Left CCACATCGTTCGGAAGCG 6250 Right GATCTGCACACACCAGTTGAG 6251 Left CTCCCAACCAAGCTCTCTT 6252 Right TGTTTTCACCAGTACGTTCCTG 6253 Left CTGCAGGAGAGGGAGCTTG 6254 Right CGACGGTCCTCCAAGTAGTT 6255 Left CATCGTTCGGAAGCGCAC 6256 Right CACACACCAGTTGAGCAGG 6257 Left GAGAAGCTCCCAACCAAG 6258 Right GATCTTGACATGCTGCGGTGTTTTC 6259 Left GCGCCACATCGTTCGGAA 6260 Right TGGGAGCCAATATTGTCTTTG 6261 Left TGCTGCAGGAGAGGGAG 6262 Right AAGCGACGGTCCTCCAAGTA 6263 Left GCCCTCCTCTTGCTGCT 6264 Right CCAGGAGGCAGCCGAAG 6265 Left CTTGTGGAGCCTCTTACA 6266 Right AGTACGTTCCTGGCTGCC 6267 Left TCTCTTGAGGATCTTGA 6268 Right CCAAAATCTGTGATCTTGACATGC 6269 Left GAAGGCGCCACATCG 6270 Right CGATCTGCACACACCAGTT 6271 Left CTCCCAACCAAGCTCT 6272 Right GATCTTGACATGCTGCGGTG 6273 Left CTGGTGGTGGCCCTGG 6274 Right TGGGAGCCAATATTGTCT EGFR Region5 276-325 bases 6275 Left ATCCTCGATGAAGCCTACGTG 6276 Right TCTCTTCCGCACCCAG 6277 Left CGATGAAGCCTACGTGATGG 6278 Right TTCTTTCTCTTCCGCAC 6279 Left GGAAATCCTCGATGAAGCCTAC 6280 Right CCAGCAGTTTGGCC 6281 Left GGAAATCCTCGATGAAGCC 6282 Right CACCCAGCAGTTTG EGFR Region5 326-375 bases 6283 Left GAAATCCTCGATGAAGCCTACG 6284 Right CCTTCTGCATGGTATTCTTTCTCTT 6285 Left GAAATCCTCGATGAAGCCTACGTGA 6286 Right CTGCATGGTATTCTTTCTCTTCCG 6287 Left AAGCAACATCTCCGAAAGCCAA 6288 Right CTCCTTCTGCATGGTATTCTTTCT 6289 Left AACATCTCCGAAAGCCAACAAG 6290 Right GCCTCCTTCTGCATGGTATT 6291 Left AGAAGCAACATCTCCGAAAG 6292 Right CATGGTATTCTTTCTCTTCCGCAC 6293 Left GGAAATCCTCGATGAAGCCT 6294 Right GCCTCCTTCTGCATGGTATTCTT 6295 Left TCGCTATCAAGGAATTAAGAGAAGC 6296 Right TCTCTTCCGCACCCAG 6297 Left CTCGATGAAGCCTACGTGATGG 6298 Right CACTTTGCCTCCTTCTGC 6299 Left TCAAGGAATTAAGAGAAGCAACATCT 6300 Right CCAGCAGTTTGGCC 6301 Left CCGTCGCTATCAAGGAATTAAGAG 6302 Right CACCCAGCAGTTTG 6303 Left GGAAATCCTCGATGAAG 6304 Right GCCTCCTTCTGCATGGT 6305 Left GGAAATCCTCGATG 6306 Right ATCCACTTGATAGGCAC 6307 Left CAACAAGGAAATCC 6308 Right CACTTTGCCTCCTTC EGFR Region5 376-425 bases 6309 Left AAGGTGAGAAAGTTAAAATTCCCGT 6310 Right CCTTCTGCATGGTATTCTTTCTCTT 6311 Left GAAAGTTAAAATTCCCGTCGCTATC 6312 Right CTCCTTCTGCATGGTATTCTTTCT 6313 Left GAAATCCTCGATGAAGCCTACG 6314 Right CTCTGGTGGGTATAGATTCTGTGTA 6315 Left CCGTCGCTATCAAGGAATTAAGAG 6316 Right CATGGTATTCTTTCTCTTCCGCAC 6317 Left GTTAAAATTCCCGTCGCTATCAAG 6318 Right TGCATGGTATTCTTTCTCTTCCG 6319 Left ATTCCCGTCGCTATCAAGGAATTA 6320 Right GCCTCCTTCTGCATGGTATTCTT 6321 Left TCGCTATCAAGGAATTAAGAGAAGC 6322 Right GCCTCCTTCTGCATGGTATT 6323 Left AACATCTCCGAAAGCCAACAAG 6324 Right ATTCCAATGCCATCCACTTGAT 6325 Left CTCGATGAAGCCTACGTGATGG 6326 Right ACTCTGGTGGGTATAGATTCTGT 6327 Left TGAGAAAGTTAAAATTCCCGTCGC 6328 Right CTTTCTCTTCCGCACCCAG 6329 Left GCAACATCTCCGAAAGCCAA 6330 Right AGATTCTGTGTAAAATTGATTCCA 6331 Left TCAAGGAATTAAGAGAAGCAACATCT 6332 Right CACTTTGCCTCCTTCTGC 6333 Left AGAAGCAACATCTCCGAAAGC 6334 Right ATTCCAATGCCATCCACTT 6335 Left ATTCCCGTCGCTATCAAGGAA 6336 Right GCCTCCTTCTGCATGGT 6337 Left TCGATGAAGCCTACGTGA 6338 Right CCCGTAGCTCCAGACATCA 6339 Left GGAAATCCTCGATGAAGCCT 6340 Right ACTCTGGTGGGTATAGATTC 6341 Left CAGAAGGTGAGAAAGTTAAAATTCC 6342 Right CCAGCAGTTTGGCC 6343 Left CCCAGAAGGTGAGAAAGTTAAAAT 6344 Right CACCCAGCAGTTTG 6345 Left TATCAAGGAATTAAGAGAAGCAACA 6346 Right TCCAATGCCATCCA 6347 Left AGAAGCAACATCTCCGAA 6348 Right ATCCACTTGATAGGCAC EGFR Region5 426-475 bases 6349 Left AAGTTAAAATTCCCGTCGCTATCA 6350 Right CTCTGGTGGGTATAGATTCTGTGTA 6351 Left TCGCTATCAAGGAATTAAGAGAAGC 6352 Right ATTCCAATGCCATCCACTTGAT 6353 Left CCGTCGCTATCAAGGAATTAAGAG 6354 Right ACTCTGGTGGGTATAGATTCTGT 6355 Left AAGGTGAGAAAGTTAAAATTCCCGT 6356 Right AGATTCTGTGTAAAATTGATTCCA 6357 Left GAAATCCTCGATGAAGCCTACG 6358 Right GTCATCAACTCCCAAACAGTCAC 6359 Left AAAGATCAAAGTGCTGGGCTC 6360 Right CATGGTATTCTTTCTCTTCCGCAC 6361 Left TCAAGGAATTAAGAGAAGCAACATCT 6362 Right CCCCGTAGCTCCAGACATC 6363 Left GGCACGGTGTATAAGGGAC 6364 Right CCTTCTGCATGGTATTCTTTCTCTT 6365 Left CCCAGAAGGTGAGAAAGTTAAA 6366 Right CCTCCTTCTGCATGGTATTCTTTC 6367 Left AGAAGCAACATCTCCGAAAGC 6368 Right CGTAGCTCCAGACATCACTCT 6369 Left CTCGATGAAGCCTACGTGATGG 6370 Right TGGATCCAAAGGTCATCAACTC 6371 Left AACATCTCCGAAAGCCAACAAG 6372 Right CATCAACTCCCAAACAGTCACCCC 6373 Left AAATCCTCGATGAAGCCTACGTGA 6374 Right AAACAGTCACCCCGTAGCTC 6375 Left AAAGATCAAAGTGCTGGGCTCCGG 6376 Right TGCATGGTATTCTTTCTCTTCCG 6377 Left GGAAATCCTCGATGAAGCCT 6378 Right AAGGTCATCAACTCCCAAACAG 6379 Left GAGAAAGTTAAAATTCCCGTCGCTA 6380 Right ATTCCAATGCCATCCACTT 6381 Left GAAGCAACATCTCCGAAAGCCAAC 6382 Right CACCCCGTAGCTCCAGAC 6383 Left TTAAAATTCCCGTCGCTATCAAGG 6384 Right ACTCTGGTGGGTATAGATTC 6385 Left CAGAAGGTGAGAAAGTTAAAATTCC 6386 Right CACTTTGCCTCCTTCTGC 6387 Left GGGCTCCGGTGCGTT 6388 Right GCCTCCTTCTGCATGGTATTCT EGFR Region6 476-525 bases 6389 Left GGAGCCTCTTACACCCAGT 6390 Right TTCTGCATGGTATTCTTTCTCTTCC 6391 Left CTCCCAACCAAGCTCTCTT 6392 Right CTCCTTCTGCATGGTATTCTTTCTC 6393 Left CTTGTGGAGCCTCTTACAC 6394 Right CATGGTATTCTTTCTCTTCCGCAC 6395 Left GAGAAGCTCCCAACCAAG 6396 Right CCTCCTTCTGCATGGTATTCTTT 6397 Left GGAGAGGGAGCTTGTGGA 6398 Right TTTCTCTTCCGCACCCAG 6399 Left CCCAGTGGAGAAGCTC 6400 Right GCCTCCTTCTGCATGGTATTC 6401 Left TCTCTTGAGGATCTTGA 6402 Right ATTCCAATGCCATCCACTTGAT 6403 Left CTCCCAACCAAGCTCT 6404 Right GCCTCCTTCTGCATGGTA 6405 Left CTGCAGGAGAGGGAGCTTG 6406 Right CCAGCAGTTTGGCC 6407 Left GAGAAGCTCCCAACC 6408 Right CACTTTGCCTCCTTCTGC 6409 Left TGCTGCAGGAGAGGGA 6410 Right CACCCAGCAGTTTG 6411 Left CCCAGTGGAGAAG 6412 Right GCCTCCTTCTGCATG EGFR Region6 526-575 bases 6413 Left CTTGTGGAGCCTCTTACACCCAG 6414 Right CCTTCTGCATGGTATTCTTTCTCTT 6415 Left CAGGAGAGGGAGCTTGTGG 6416 Right CTGCATGGTATTCTTTCTCTTCCG 6417 Left CTGCAGGAGAGGGAGCTTG 6418 Right CCTCCTTCTGCATGGTATTCTTTC 6419 Left CTTGTGGAGCCTCTTACAC 6420 Right CATGGTATTCTTTCTCTTCCGCAC 6421 Left CTCCCAACCAAGCTCTCTT 6422 Right ATTCCAATGCCATCCACTTGAT 6423 Left GAGAAGCTCCCAACCAAG 6424 Right GCCTCCTTCTGCATGGTATTCT 6425 Left TGCAGGAGAGGGAGC 6426 Right CTTTCTCTTCCGCACCCAG 6427 Left CCCAGTGGAGAAGCTC 6428 Right GCCTCCTTCTGCATGGTAT 6429 Left TCTCTTGAGGATCTTGA 6430 Right AGATTCTGTGTAAAATTGATTCCA 6431 Left CTCCCAACCAAGCTCT 6432 Right CACTTTGCCTCCTTCTGC 6433 Left CCACATCGTTCGGAAGCG 6434 Right CCAGCAGTTTGGCC 6435 Left GAGAAGCTCCCAACC 6436 Right ATTCCAATGCCATCCACTT 6437 Left CATCGTTCGGAAGCGCAC 6438 Right CACCCAGCAGTTTG 6439 Left CTTGTGGAGCCTCTTA 6440 Right GCCTCCTTCTGCATGG 6441 Left CCCAGTGGAGAAG 6442 Right ATCCACTTGATAGGCAC 6443 Left CTTGTGGAGCCTC 6444 Right CACTTTGCCTCCTTC EGFR Region6 576-625 bases 6445 Left CTTGTGGAGCCTCTTACACCCAG 6446 Right CTCTGGTGGGTATAGATTCTGTGTA 6447 Left CCACATCGTTCGGAAGCG 6448 Right CCTTCTGCATGGTATTCTTTCTCTT 6449 Left ATCGGCCTCTTCATGCGAA 6450 Right CATGGTATTCTTTCTCTTCCGCAC 6451 Left GAAGGCGCCACATCGTTC 6452 Right CCTCCTTCTGCATGGTATTCTTTC 6453 Left CTTGTGGAGCCTCTTACACC 6454 Right ACTCTGGTGGGTATAGATTCTGT 6455 Left CAGGAGAGGGAGCTTGTGG 6456 Right ATTCCAATGCCATCCACTTGAT 6457 Left CGTTCGGAAGCGCACG 6458 Right CTGCATGGTATTCTTTCTCTTCCG 6459 Left CTGCAGGAGAGGGAGCTTG 6460 Right GCCTCCTTCTGCATGGTATTCT 6461 Left CTCCCAACCAAGCTCTCTT 6462 Right CGTAGCTCCAGACATCACTCT 6463 Left GGATCGGCCTCTTCATGC 6464 Right CTTTCTCTTCCGCACCCAG 6465 Left GAGAAGCTCCCAACCAAG 6466 Right AGATTCTGTGTAAAATTGATTCCA 6467 Left GCGCCACATCGTTCGGAA 6468 Right GCCTCCTTCTGCATGGTAT 6469 Left CGAAGGCGCCACATCG 6470 Right CACTTTGCCTCCTTCTGC 6471 Left TGCTGCAGGAGAGGGAG 6472 Right ATTCCAATGCCATCCACTT 6473 Left CCCAGTGGAGAAGCTC 6474 Right CCCGTAGCTCCAGACATCAC 6475 Left TCTCTTGAGGATCTTGA 6476 Right AAAGGTCATCAACTCCCAAACAG 6477 Left CTCCCAACCAAGCTCT 6478 Right AAACAGTCACCCCGTAGCTC 6479 Left GAGAAGCTCCCAACC 6480 Right ACCCCGTAGCTCCAGACAT 6481 Left CTTGTGGAGCCTCTTAC 6482 Right ACTCTGGTGGGTATAGATTC 6483 Left CTCTTGCTGCTGGTGGTG 6484 Right CCAGCAGTTTGGCC EGFR Region6 750-1250 bases 6485 Left TTTGTGGAGAACTCTGAGTGCATA 6486 Right CTCTGGTGGGTATAGATTCTGTGTA 6487 Left CAGGAGTCATGGGAGAAAACAAC 6488 Right CCTTCTGCATGGTATTCTTTCTCTT 6489 Left ACCAAAATTATAAGCAACAGAGGTG 6490 Right CCTCCTTCTGCATGGTATTCTTT 6491 Left TATAAGCAACAGAGGTGAAAACAGC 6492 Right ACTCTGGTGGGTATAGATTCTGT 6493 Left GAGTCATGGGAGAAAACAACACC 6494 Right TTTTGGAGAATTCGATGATCAACTC 6495 Left GAGTTTGTGGAGAACTCTGAGTG 6496 Right CATGGTATTCTTTCTCTTCCGCAC 6497 Left GAGAAAACAACACCCTGGTCTG 6498 Right TGCATGGTATTCTTTCTCTTCCG 6499 Left AAAACAACACCCTGGTCTGGAAGTA 6500 Right GATTCCGTCATATGGCTTGGATC 6501 Left GCAGGAGTCATGGGAGAAAAC 6502 Right CTATCATCCAGCACTTGACCATG 6503 Left CCAAGGGAGTTTGTGGAGAAC 6504 Right AAAGGTCATCAACTCCCAAACAG 6505 Left GAGAAAACAACACCCTGGTCTGGAA 6506 Right CGACTATCTGCGTCTATCATCCAG 6507 Left AACGAATGGGCCTAAGATCCC 6508 Right CATTTCTATCAATGCAAGCCACG 6509 Left GTCAGAAAACCAAAATTATAAGCAA CAG 6510 Right ATTCCAATGCCATCCACTTGAT 6511 Left ATCCAAACTGCACCTACGGAT 6512 Right GATCCAAAGGTCATCAACTCCCAA 6513 Left CCATGAACATCACCTGCACAG 6514 Right CTATCTGCGTCTATCATCCAGCAC 6515 Left AATTATAAGCAACAGAGGTGAAAAC 6516 Right CCGTAGCTCCAGACATCACT 6517 Left AGGTGAAAACAGCTGCAAGG 6518 Right GTCATCAACTCCCAAACAGTCAC 6519 Left GGGAGTTTGTGGAGAACTCTG 6520 Right CATCCAGCACTTGACCATGATC 6521 Left CCATCCAAACTGCACCTACG 6522 Right TGATCAACTCACGGAACTTTGG 6523 Left AATGGGCCTAAGATCCCGTC 6524 Right CGTCTATCATCCAGCACTTGAC

TABLE 13 BRAF cDNA Capture Primer List for NGS Panel Seq. ID Primer Sequence BRAF Region1 75-125 bases 6525 Left gacaggaatcgaatgaaaacacttg 6526 Right tttcccttgtagactgttccaaat 6527 Left ggaatcgaatgaaaacacttggtag 6528 Right cactttcccttgtagactgttcc 6529 Left ggaatcgaatgaaaacac 6530 Right ccactttcccttgtagactgt 6531 Left ggaatcgaatgaaaa 6532 Right ccactttcccttgtagac BRAF Region1 126-175 bases 6533 Left gaagacaggaatcgaatgaaaacac 6534 Right gctgtcacattcaacattttcactg 6535 Left aggaatcgaatgaaaacacttggta 6536 Right cacattcaacattttcactgccac 6537 Left tcttcatcctcagaagacaggaatc 6538 Right caacattttcactgccacatcac 6539 Left aagtcatcttcatcctcagaagaca 6540 Right cattttcactgccacatcaccat 6541 Left agacaggaatcgaatgaaaacacttg 6542 Right tttcccttgtagactgttccaaat 6543 Left ctcagaagacaggaatcgaatgaaa 6544 Right catcaccatgccactttccc 6545 Left aatcgaatgaaaacacttggtagac 6546 Right gctgtcacattcaacattttca 6547 Left aaatctccaggacctcagcgagaaa 6548 Right actttcccttgtagactgttcca 6549 Left atcctcagaagacaggaatcgaa 6550 Right atcaccatgccactttcccttg 6551 Left cagcgagaaaggaagtcatct 6552 Right gccactttcccttgtagactgtt 6553 Left aagtcatcttcatcctcagaag 6554 Right atcaccatgccactttcccttgtag 6555 Left ctccaggacctcagcgag 6556 Right tgccactttcccttgtagact 6557 Left catcttcatcctcagaagacagga 6558 Right catcaccatgccacttt 6559 Left ctcagcgagaaaggaagtca 6560 Right catcaccatgccac BRAF Region1 176-225 bases 6561 Left aagtcatcttcatcctcagaagaca 6562 Right gctgtcacattcaacattttcactg 6563 Left actaactaacgtgaaagccttacag 6564 Right attttcactgccacatcaccat 6565 Left ccttacagaaatctccaggacct 6566 Right tcacattcaacattttcactgcca 6567 Left aagccttacagaaatctccagga 6568 Right caacattttcactgccacatcac 6569 Left ctcagcgagaaaggaagtcatct 6570 Right cttgtaactgctgaggtgtaggt 6571 Left ttacagaaatctccaggacctcagc 6572 Right tgctgtcacattcaacattttca 6573 Left ccaggacctcagcgagaaa 6574 Right ctgaggtgtaggtgctgtca 6575 Left ctaacgtgaaagccttacagaaat 6576 Right caacattttcactgccacat 6577 Left aagtcatcttcatcctcagaag 6578 Right ctgctgaggtgtaggtgct 6579 Left tcagcgagaaaggaagtca 6580 Right gcttgtaactgctgaggtgta 6581 Left gaaatctccaggacctcagcgag 6582 Right tgctgtcacattcaacattt 6583 Left aagtcatcttcatcctcag 6584 Right ttttgaaggcttgtaactgctg 6585 Left ctcactaactaacgtgaaagcctta 6586 Right catcaccatgcca 6587 Left aagtcatcttcatcct 6588 Right aggcttgtaactgctgaggt 6589 Left ctcagcgagaaaggaag 6590 Right tgctgtcacattcaaca 6591 Left agccttacagaaatctcca 6592 Right gctgtcacattca BRAF Region1 226-275 bases 6593 Left agtcatcttcatcctcagaagaca 6594 Right tgaagagtaggatattcacatgtcg 6595 Left ctcactaactaacgtgaaagcctta 6596 Right cacattcaacattttcactgccac 6597 Left actaactaacgtgaaagccttacag 6598 Right ttttgaaggcttgtaactgctgag 6599 Left cctcattacctggctcactaacta 6600 Right gctgtcacattcaacattttcactg 6601 Left actaacgtgaaagccttacagaaat 6602 Right cttgtaactgctgaggtgtaggt 6603 Left attacctggctcactaactaacgt 6604 Right caacattttcactgccacatcac 6605 Left ctggctcactaactaacgtgaaag 6606 Right tgctgtcacattcaacattttca 6607 Left aacgtgaaagccttacagaaatctc 6608 Right ggcttgtaactgctgaggtgta 6609 Left cttacagaaatctccaggacctca 6610 Right taactgctgaggtgtaggtgct 6611 Left aagccttacagaaatctccagga 6612 Right ttttgaaggcttgtaactgct 6613 Left cctcattacctggctcactaa 6614 Right cattttcactgccacatcaccat 6615 Left gaaatctccaggacctcagcgagaa 6616 Right ctgaggtgtaggtgctgtca 6617 Left agtcatcttcatcctcagaag 6618 Right gcccatgaagagtaggatattc 6619 Left ctcagcgagaaaggaagtcatct 6620 Right catgtcgtgttttcctga 6621 Left aaatctccaggacctcagcg 6622 Right gttttcctgagtactcctac 6623 Left cccctgcctcattacctg 6624 Right tgctgtcacattcaacattt 6625 Left aagtcatcttcatcctcag 6626 Right tgaagagtaggatattcacatg 6627 Left ctcagcgagaaaggaagtca 6628 Right tttgaaggcttgtaact 6629 Left cctcattacctggctcac 6630 Right tgctgtcacattcaaca 6631 Left caggtttgtctgctacccc 6632 Right catcaccatgcca BRAF Region1 276-325 bases 6633 Left aagtcatcttcatcctcagaagaca 6634 Right ggaatagcccatgaagagtaggata 6635 Left actaactaacgtgaaagccttacag 6636 Right tgaagagtaggatattcacatgtcg 6637 Left ctcattacctggctcactaactaac 6638 Right ttttgaaggcttgtaactgctgag 6639 Left aacgtgaaagccttacagaaatctc 6640 Right atagcccatgaagagtaggatattc 6641 Left ttgatgacttgattagagaccaagg 6642 Right attttcactgccacatcaccat 6643 Left gatttcgtggtgatggaggatc 6644 Right gctgtcacattcaacattttcactg 6645 Left attacctggctcactaactaacgtg 6646 Right gcttgtaactgctgaggtgtag 6647 Left caaggatttcgtggtgatggag 6648 Right tcacattcaacattttcactgcca 6649 Left cctcattacctggctcactaact 6650 Right cttgtaactgctgaggtgtaggtg 6651 Left ctggctcactaactaacgtgaaag 6652 Right ttgaaggcttgtaactgctgaggtg 6653 Left aagccttacagaaatctccagga 6654 Right ggaatagcccatgaagagtagg 6655 Left tattgatgacttgattagagacca 6656 Right caacattttcactgccacatcac 6657 Left ttgattagagaccaaggatttcgtg 6658 Right tgctgtcacattcaacatttt 6659 Left gctcactaactaacgtgaaagcctt 6660 Right ttttgaaggcttgtaactgct 6661 Left cttacagaaatctccaggacctca 6662 Right tgaagagtaggatattcacatg 6663 Left cccctgcctcattacctgg 6664 Right gtaactgctgaggtgtaggtgctg 6665 Left aaatctccaggacctcagcgagaaa 6666 Right cagttgtggctttgtggaat 6667 Left ctcagcgagaaaggaagtcatct 6668 Right ggtaacaatagccagttgtg 6669 Left atgacttgattagagaccaaggatt 6670 Right aacattttcactgccacat 6671 Left ctaacgtgaaagccttacagaaat 6672 Right catgtcgtgttttcctgag BRAF Region1 326-375 bases 6673 Left ctcattacctggctcactaactaac 6674 Right ggaatagcccatgaagagtaggata 6675 Left ctcactaactaacgtgaaagcctta 6676 Right tgaagagtaggatattcacatgtcg 6677 Left agtcatcttcatcctcagaagaca 6678 Right gatatggagatggtgatacaagctg 6679 Left acttgattagagaccaaggatttcg 6680 Right ttttgaaggcttgtaactgctgag 6681 Left aacgtgaaagccttacagaaatctc 6682 Right gaatagcccatgaagagtaggatattc 6683 Left tgatgacttgattagagaccaagga 6684 Right cttgtaactgctgaggtgtaggt 6685 Left actaacgtgaaagccttacagaaat 6686 Right ggaatagcccatgaagagtagg 6687 Left tataaacacaatagaacctgtcaat 6688 Right gctgtcacattcaacattttcactg 6689 Left ctcagcgagaaaggaagtcatct 6690 Right atggagatggtgatacaagctggag 6691 Left gcatataaacacaatagaacctgtc 6692 Right cattttcactgccacatcacca 6693 Left tattgatgacttgattagagacca 6694 Right cacattcaacattttcactgccac 6695 Left gatttcgtggtgatggaggatc 6696 Right gaaggcttgtaactgctgaggtgta 6697 Left aaacacaatagaacctgtcaatatt 6698 Right tgctgtcacattcaacattttca 6699 Left caaggatttcgtggtgatggag 6700 Right taactgctgaggtgtaggtgct 6701 Left attacctggctcactaactaacgtg 6702 Right cagttgtggctttgtggaat 6703 Left actaactaacgtgaaagccttacag 6704 Right ggtaacaatagccagttgtg 6705 Left cttacagaaatctccaggacctca 6706 Right agccctcacaccactgg 6707 Left ccgatcctcatcagctccc 6708 Right caacattttcactgccacatca 6709 Left ctggctcactaactaacgtgaaag 6710 Right tgaagagtaggatattcacatg 6711 Left ccaggacctcagcgagaaa 6712 Right tgatatggagatggtgatacaag BRAF Region1 376-425 bases 6713 Left actaactaacgtgaaagccttacag 6714 Right gatatggagatggtgatacaagctg 6715 Left cttgattagagaccaaggatttcgt 6716 Right ggaatagcccatgaagagtaggata 6717 Left aagtcatcttcatcctcagaagaca 6718 Right gcagtctgtcgtgcaatatctataa 6719 Left gaagatcatcgaaatcaatttgggc 6720 Right gctgtcacattcaacattttcactg 6721 Left ttgatgacttgattagagaccaagg 6722 Right tgaagagtaggatattcacatgtcg 6723 Left tgacttgattagagaccaaggattt 6724 Right atagcccatgaagagtaggatattc 6725 Left gatcatcgaaatcaatttgggcaac 6726 Right cacattcaacattttcactgccac 6727 Left ctcactaactaacgtgaaagcctta 6728 Right atggagatggtgatacaagctggag 6729 Left gcatataaacacaatagaacctgtc 6730 Right ttttgaaggcttgtaactgctgag 6731 Left cttccgaccagcagatgaagatc 6732 Right caacattttcactgccacatcac 6733 Left ctcagcgagaaaggaagtcatct 6734 Right gcagtctgtcgtgcaatatcta 6735 Left cctcattacctggctcactaacta 6736 Right tgatatggagatggtgatacaag 6737 Left ttacctggctcactaactaacgt 6738 Right ttggtctcaatgatatggagatg 6739 Left gatttcgtggtgatggaggatc 6740 Right ggaatagcccatgaagagtagg 6741 Left ataaacacaatagaacctgtcaata 6742 Right cttgtaactgctgaggtgtaggt 6743 Left agaaatctccaggacctcagc 6744 Right cgtgcaatatctataagtttgatca 6745 Left actaacgtgaaagccttacagaaat 6746 Right ctggagccctcacaccac 6747 Left aatctccaggacctcagcgagaaag 6748 Right ctgtcgtgcaatatctataagtttg 6749 Left gagaccgatcctcatcagctc 6750 Right aacattttcactgccacatcaccat 6751 Left gaaatctccaggacctcagcgaga 6752 Right tgatcatctcaaatttggtctcaa BRAF Region1 426-475 bases 6753 Left actaacgtgaaagccttacagaaat 6754 Right gcagtctgtcgtgcaatatctataa 6755 Left ccttcaaaatccattccaattccac 6756 Right gctgtcacattcaacattttcactg 6757 Left attgatgacttgattagagaccaagg 6758 Right ggaatagcccatgaagagtaggata 6759 Left actaactaacgtgaaagccttacag 6760 Right agtctgtcgtgcaatatctataagtt 6761 Left gaagatcatcgaaatcaatttgggc 6762 Right ttttgaaggcttgtaactgctga 6763 Left tgcatataaacacaatagaacctgtc 6764 Right tgaagagtaggatattcacatgtcg 6765 Left caaaatccattccaattccacagc 6766 Right cacattcaacattttcactgccac 6767 Left aacgtgaaagccttacagaaatctc 6768 Right gtgtaagtaatccatgccctgtg 6769 Left gatttcgtggtgatggaggatc 6770 Right gatatggagatggtgatacaagctg 6771 Left ctcattacctggctcactaactaac 6772 Right cgtgcaatatctataagtttgatca 6773 Left aagtcatcttcatcctcagaagaca 6774 Right ggatgattgacttggcgtgtaag 6775 Left gatcatcgaaatcaatttgggcaac 6776 Right cttgtaactgctgaggtgtaggt 6777 Left ctcactaactaacgtgaaagcctta 6778 Right gcagtctgtcgtgcaatatcta 6779 Left gtccgtctccttcaaaatccatt 6780 Right caacattttcactgccacatcac 6781 Left attacctggctcactaactaacgtg 6782 Right tgatcatctcaaatttggtctcaa 6783 Left atattgatgacttgattagagacca 6784 Right aatagcccatgaagagtaggatattc 6785 Left ctcagcgagaaaggaagtcatc 6786 Right gaggtctctgtggatgattgactt 6787 Left ttacagaaatctccaggacctca 6788 Right cttggcgtgtaagtaatccatgc 6789 Left caaattctcaccagtccgtctc 6790 Right aacattttcactgccacatcaccat 6791 Left ctggctcactaactaacgtgaaa 6792 Right cgtgcaatatctataagtttgatcatct BRAF Region1 476-525 bases 6793 Left cttgattagagaccaaggatttcgt 6794 Right gatatggagatggtgatacaagctg 6795 Left tcactaactaacgtgaaagccttac 6796 Right ttattactcttgaggtctctgtgga 6797 Left gaagatcatcgaaatcaatttgggc 6798 Right ggaatagcccatgaagagtaggata 6799 Left aagtcatcttcatcctcagaagaca 6800 Right actgtagctagaccaaaatcaccta 6801 Left gatcatcgaaatcaatttgggcaac 6802 Right tgaagagtaggatattcacatgtcg 6803 Left ctcattacctggctcactaactaac 6804 Right ctcttgaggtctctgtggatgatt 6805 Left actaacgtgaaagccttacagaaat 6806 Right gtctctgtggatgattgacttgg 6807 Left cttcaaaatccattccaattccaca 6808 Right ttttgaaggcttgtaactgctgag 6809 Left ctaactaacgtgaaagccttacaga 6810 Right gaggtctctgtggatgattgact 6811 Left gatttcgtggtgatggaggatc 6812 Right gcagtctgtcgtgcaatatctataa 6813 Left tgatgacttgattagagaccaagga 6814 Right atggagatggtgatacaagctggag 6815 Left caaattctcaccagtccgtctc 6816 Right gctgtcacattcaacattttcactg 6817 Left ctggctcactaactaacgtgaaag 6818 Right gtgtaagtaatccatgccctgtg 6819 Left cttacagaaatctccaggacctca 6820 Right ggatgattgacttggcgtgtaag 6821 Left gtccgtctccttcaaaatccattc 6822 Right cttgtaactgctgaggtgtaggt 6823 Left aaattctcaccagtccgtctccttc 6824 Right cacattcaacattttcactgccac 6825 Left cttccgaccagcagatgaagatc 6826 Right atagcccatgaagagtaggatattc 6827 Left ttacctggctcactaactaacgtga 6828 Right cttggcgtgtaagtaatccatgc 6829 Left caaggatttcgtggtgatggag 6830 Right ctgtcgtgcaatatctataagtttg 6831 Left gactgccctaacatctggatcat 6832 Right attttcactgccacatcaccat BRAF Region2 75-125 bases 6833 Left aatcatccacagagacctcaagag 6834 Right tgttcaaactgatgggaccc 6835 Left tccacagagacctcaagagtaa 6836 Right agacaactgttcaaactga 6837 Left tcaatcatccacagagacctcaa 6838 Right tgttcaaactgatggga 6839 Left caagtcaatcatccacagagacc 6840 Right tgggacccactc BRAF Region2 126-175 bases 6841 Left aatcatccacagagacctcaagag 6842 Right attctgatgacttctggtgccat 6843 Left caagtcaatcatccacagagacc 6844 Right aactgttcaaactgatgggacc 6845 Left catccacagagacctcaagagtaa 6846 Right cacaaaatggatccagacaact 6847 Left tcaatcatccacagagacctcaa 6848 Right attctgatgacttctggtgc 6849 Left cacagggcatggattacttacac 6850 Right agacaactgttcaaactgat 6851 Left cttacacgccaagtcaatcatcc 6852 Right aactgttcaaactgatggg 6853 Left gcatggattacttacacgccaag 6854 Right cacaaaatggatccagaca 6855 Left gccaagtcaatcatccacagag 6856 Right tgccatccacaaaatg 6857 Left ttacacgccaagtcaatcatccaca 6858 Right agacaactgttcaaact 6859 Left ggcatggattacttacacgcc 6860 Right cacaaaatggatccag 6861 Left ttatagatattgcacgacagactgc 6862 Right tgggacccactc 6863 Left cttacacgccaagtcaatca 6864 Right tgccatccacaaa 6865 Left gcacgacagactgcacag 6866 Right agacaactgttcaa 6867 Left acgccaagtcaa 6868 Right ctgatgacttctgg BRAF Region2 176-225 bases 6869 Left ttatagatattgcacgacagactgc 6870 Right attctgatgacttctggtgccat 6871 Left cttacacgccaagtcaatcatcc 6872 Right tctgactgaaagctgtatggatttt 6873 Left tacacgccaagtcaatcatccacag 6874 Right atgcatatacatctgactgaaagct 6875 Left aacttatagatattgcacgacagac 6876 Right cacaaaatggatccagacaact 6877 Left catggattacttacacgccaag 6878 Right tctgactgaaagctgtatggat 6879 Left cacagggcatggattacttacac 6880 Right attctgatgacttctggtgc 6881 Left aaacttatagatattgcacgaca 6882 Right cacaaaatggatccagaca 6883 Left acacgccaagtcaatca 6884 Right aatgcatatacatctgactgaaa 6885 Left tgatcaaacttatagatattgcacg 6886 Right tgccatccacaaaatg 6887 Left gcatggattacttacacgcc 6888 Right ctgactgaaagctgtatg 6889 Left gcacgacagactgcacag 6890 Right ttttatcttgcattctgat 6891 Left gagatgatcaaacttatagatattgc 6892 Right agacaactgttcaaact 6893 Left tgagaccaaatttgagatgatc 6894 Right cacaaaatggatccag 6895 Left cacagggcatggattactt 6896 Right attctgatgacttctgg 6897 Left acacgccaagtcaa 6898 Right aatgcatatacatctgactg 6899 Left catctccatatcattgagacca 6900 Right agacaactgttcaa 6901 Left gagatgatcaaacttatagatat 6902 Right tgccatccacaaa 6903 Left cacagggcatggatta 6904 Right ttttatcttgcattct BRAF Region2 226-275 bases 6905 Left ttatagatattgcacgacagactgc 6906 Right tccaaatgcatatacatctgactga 6907 Left aacttatagatattgcacgacagac 6908 Right tctgactgaaagctgtatggatttt 6909 Left tcaaacttatagatattgcacgaca 6910 Right atgcatatacatctgactgaaagct 6911 Left atcaccatctccatatcattgagac 6912 Right attctgatgacttctggtgccat 6913 Left cacagggcatggattacttacac 6914 Right acagaacaattccaaatgcatataca 6915 Left gcatggattacttacacgccaa 6916 Right acaattccaaatgcatatacatctgac 6917 Left tgatcaaacttatagatattgcacg 6918 Right tctgactgaaagctgtatggat 6919 Left cagcttgtatcaccatctccatatc 6920 Right ttctgatgacttctggtgc 6921 Left gcacgacagactgcacag 6922 Right acagaacaattccaaatgcatat 6923 Left cacagggcatggattactta 6924 Right tccagtcatcaattcatacaga 6925 Left gagatgatcaaacttatagatattgc 6926 Right tctgactgaaagctgtatg 6927 Left tgtatcaccatctccatatcattga 6928 Right tgccatccacaaaatg 6929 Left cacagggcatggattac 6930 Right caattccaaatgcatatacatct 6931 Left ccagcttgtatcaccatctccat 6932 Right cacaaaatggatccag 6933 Left ttgagaccaaatttgagatgatc 6934 Right attctgatgacttctgg 6935 Left ctccagcttgtatcaccatctc 6936 Right tgccatccacaaa 6937 Left cacagggcatggat 6938 Right acagaacaattccaaatgca 6939 Left gagatgatcaaacttatagatat 6940 Right tttatcttgcattctga BRAF Region2 276-325 bases 6941 Left cagcttgtatcaccatctccatatc 6942 Right tctgactgaaagctgtatggatttt 6943 Left atcaccatctccatatcattgagac 6944 Right atgcatatacatctgactgaaagct 6945 Left acttatagatattgcacgacagactg 6946 Right ctgtccagtcatcaattcatacaga 6947 Left cacagggcatggattacttacac 6948 Right aaaattatctggtccctgttgttga 6949 Left caaacttatagatattgcacgacaga 6950 Right ccaaatgcatatacatctgactgaa 6951 Left gcatggattacttacacgccaa 6952 Right attatctggtccctgttgttgatgt 6953 Left cacaactggctattgttacccag 6954 Right attctgatgacttctggtgccat 6955 Left gcttgtatcaccatctccatatcatt 6956 Right tctgactgaaagctgtatggat 6957 Left gcacgacagactgcacag 6958 Right cctgttgttgatgtttgaataaggt 6959 Left gagatgatcaaacttatagatattgc 6960 Right attccaaatgcatatacatctgact 6961 Left ttgagaccaaatttgagatgatc 6962 Right acagaacaattccaaatgcatataca 6963 Left cacagggcatggattactta 6964 Right ggtccctgttgttgatgtttgaa 6965 Left tgatcaaacttatagatattgcacgac 6966 Right ctgtccagtcatcaattcata 6967 Left ccacaactggctattgttacc 6968 Right attctgatgacttctggtgc 6969 Left ttgagaccaaatttgagatg 6970 Right acaattccaaatgcatatacatctg 6971 Left ctccagcttgtatcaccatctcc 6972 Right tctgactgaaagctgtatg 6973 Left gagatgatcaaacttatagat 6974 Right acagaacaattccaaatgcatat 6975 Left cacagggcatggattac 6976 Right aaaattatctggtccctgttgt 6977 Left tatcctactatcatgggctattcc 6978 Right cacaaaatggatccag 6979 Left attccacaaagccacaactg 6980 Right tgccatccacaaaatg BRAF Region2 326-375 bases 6981 Left ttatagatattgcacgacagactgc 6982 Right aaaattatctggtccctgttgttga 6983 Left atcaccatctccatatcattgagac 6984 Right ctgtccagtcatcaattcatacaga 6985 Left ccagcttgtatcaccatctccatat 6986 Right tccaaatgcatatacatctgactga 6987 Left cacaactggctattgttacccag 6988 Right tctgactgaaagctgtatggatttt 6989 Left tatcctactcttcatgggctattcc 6990 Right attctgatgacttctggtgccat 6991 Left aacttatagatattgcacgacagac 6992 Right tatctggtccctgttgttgatgttt 6993 Left gcatggattacttacacgccaa 6994 Right ttttggacagttactccgtacctta 6995 Left cacagggcatggattacttacac 6996 Right ccgtaccttactgagatctggag 6997 Left tcaaacttatagatattgcacgaca 6998 Right ggtatcctcgtcccaccataaaa 6999 Left ctccagcttgtatcaccatctcc 7000 Right acagaacaattccaaatgcatataca 7001 Left gctattccacaaagccacaac 7002 Right aatgcatatacatctgactgaaagc 7003 Left agatgatcaaacttatagatattgcac 7004 Right ggtccctgttgttgatgtttgaa 7005 Left gcttgtatcaccatctccatatcatt 7006 Right ctgtccagtcatcaattcatac 7007 Left ccacaactggctattgttacc 7008 Right acaattccaaatgcatatacatctgac 7009 Left tgaatatcctactcttcatgggcta 7010 Right attctgatgacttctggtgc 7011 Left agatgatcaaacttatagatattg 7012 Right cctgttgttgatgtttgaataaggt 7013 Left gcacgacagactgcacag 7014 Right aggtatcctcgtcccaccata 7015 Left ttgagaccaaatttgagatgatc 7016 Right aaaattatctggtccctgttgt 7017 Left ccagtggtgtgagggctc 7018 Right tctgactgaaagctgtatggat 7019 Left cacagggcatggattactta 7020 Right caggtatcctcgtcccacc BRAF Region2 376-425 bases 7021 Left cagcttgtatcaccatctccatatc 7022 Right aaaattatctggtccctgttgttga 7023 Left tgaatatcctactcttcatgggcta 7024 Right tctgactgaaagctgtatggatttt 7025 Left tatcctactcttcatgggctattcc 7026 Right ctgtccagtcatcaattcatacaga 7027 Left cgacatgtgaatatcctactcttca 7028 Right atgcatatacatctgactgaaagct 7029 Left acttatagatattgcacgacagact 7030 Right ttttggacagttactccgtacctta 7031 Left atcaccatctccatatcattgagac 7032 Right ggtccctgttgttgatgtttgaa 7033 Left ccacaactggctattgttaccc 7034 Right cctgttgttgatgtttgaataaggt 7035 Left tcaaacttatagatattgcacgaca 7036 Right cttttggacagttactccgtacc 7037 Left ctccagcttgtatcaccatctcc 7038 Right attatctggtccctgttgttgatgt 7039 Left acgacatgtgaatatcctactct 7040 Right tccaaatgcatatacatctgactga 7041 Left tcagcagttacaagccttcaaaa 7042 Right ttctgatgacttctggtgccat 7043 Left tgatcaaacttatagatattgcacg 7044 Right ccgtaccttactgagatctggag 7045 Left cacagggcatggattacttacac 7046 Right ggcttttggacagttactccg 7047 Left gcttgtatcaccatctccatatcatt 7048 Right aaaattatctggtccctgttgt 7049 Left gagatgatcaaacttatagatattgc 7050 Right ggtatcctcgtcccaccataaaa 7051 Left tgaatatcctactcttcatggg 7052 Right acagaacaattccaaatgcatataca 7053 Left ttgagaccaaatttgagatgatc 7054 Right aggtatcctcgtcccaccata 7055 Left acctacacctcagcagttacaag 7056 Right attctgatgacttctggtgc 7057 Left attccacaaagccacaactg 7058 Right attccaaatgcatatacatctgac 7059 Left gcacgacagactgcacag 7060 Right cactctgccattaatctcttcat BRAF Region2 426-475 bases 7061 Left tatcctactcttcatgggctattcc 7062 Right aaaattatctggtccctgttgttga 7063 Left tgaatatcctactcttcatgggcta 7064 Right ctgtccagtcatcaattcatacaga 7065 Left agcttgtatcaccatctccatatca 7066 Right ttttggacagttactccgtacctta 7067 Left cgacatgtgaatatcctactcttca 7068 Right attatctggtccctgttgttgatgt 7069 Left atcaccatctccatatcattgagac 7070 Right cttttggacagttactccgtacc 7071 Left ctcagcagttacaagccttcaaaa 7072 Right tccaaatgcatatacatctgactga 7073 Left acctacacctcagcagttacaag 7074 Right atgcatatacatctgactgaaagct 7075 Left tatagatattgcacgacagactgc 7076 Right gagaatttggggaaagagtggtc 7077 Left gatgtggcagtgaaaatgttgaatg 7078 Right attctgatgacttctggtgccat 7079 Left cagggcatggattacttacacg 7080 Right agtggtctctcatctcttttctttt 7081 Left tgtatcaccatctccatatcattga 7082 Right ccgtaccttactgagatctggag 7083 Left tgacagcacctacacctcag 7084 Right tctgactgaaagctgtatggatttt 7085 Left cacaactggctattgttacccag 7086 Right ggtatcctcgtcccaccataaaa 7087 Left gcatggattacttacacgccaa 7088 Right aatagaggcgagaatttggggaaag 7089 Left tacacctcagcagttacaagcctt 7090 Right acagaacaattccaaatgcatataca 7091 Left aacttatagatattgcacgacagac 7092 Right cactctgccattaatctcttcat 7093 Left acgacatgtgaatatcctactct 7094 Right ggtccctgttgttgatgtttgaa 7095 Left taggagtactcaggaaaacacgac 7096 Right acagaacaattccaaatgcatat 7097 Left ctccagcttgtatcaccatctcc 7098 Right ggcttttggacagttactccg 7099 Left tcaaacttatagatattgcacgaca 7100 Right agtggtctctcatctcttttct BRAF Region2 476-525 bases 7101 Left cgacatgtgaatatcctactcttca 7102 Right aaaattatctggtccctgttgttga 7103 Left cagtgaaaatgttgaatgtgacagc 7104 Right ctgtccagtcatcaattcatacaga 7105 Left tatcctactcttcatgggctattcc 7106 Right tttggacagttactccgtacctta 7107 Left gcttgtatcaccatctccatatcatt 7108 Right agtggtctctcatctcttttctttt 7109 Left ttatagatattgcacgacagactgc 7110 Right aatagaggcgagaatttggggaaag 7111 Left gatgtggcagtgaaaatgttgaatg 7112 Right ccaaatgcatatacatctgactgaa 7113 Left atcaccatctccatatcattgagac 7114 Right gagaatttggggaaagagtggtc 7115 Left tgaatatcctactcttcatgggcta 7116 Right ggtatcctcgtcccaccataaaa 7117 Left catggtgatgtggcagtgaaaat 7118 Right tctgactgaaagctgtatggatttt 7119 Left tcagcagttacaagccttcaaaa 7120 Right attatctggtccctgttgttgatgt 7121 Left gtctacaagggaaagtggcatg 7122 Right atgcatatacatctgactgaaagct 7123 Left aacttatagatattgcacgacagac 7124 Right aatagaggcgagaatttgggga 7125 Left ccacaactggctattgttaccc 7126 Right cttttggacagttactccgtacc 7127 Left acctacacctcagcagttaca 7128 Right cctgttgttgatgtttgaataaggt 7129 Left ccagcttgtatcaccatctccatat 7130 Right cactctgccattaatctcttcat 7131 Left tggaacagtctacaagggaaagt 7132 Right ttctgatgacttctggtgccat 7133 Left aacagtctacaagggaaagtggc 7134 Right attccaaatgcatatacatctgact 7135 Left ctacacctcagcagttacaagcc 7136 Right ggtccctgttgttgatgtttgaa 7137 Left tgacagcacctacacctcag 7138 Right acagaacaattccaaatgcatataca 7139 Left cacagggcatggattacttacac 7140 Right aaggagggttctgatgcactg BRAF Region2 750-1250 bases 7141 Left taaatttcaccagcgttgtagtaca 7142 Right aaaattatctggtccctgttgttga 7143 Left catgtggttataaatttcaccagcg 7144 Right ctgtccagtcatcaattcatacaga 7145 Left cttgattagagaccaaggatttcgt 7146 Right ttttggacagttactccgtacctta 7147 Left tatgaccaacttgatttgctgtttg 7148 Right cctgttgttgatgtttgaataaggt 7149 Left gaagatcatcgaaatcaatttgggc 7150 Right agtggtctctcatctcttttctttt 7151 Left ccttcaaaatccattccaattccac 7152 Right attatctggtccctgttgttgatgt 7153 Left tgatgacttgattagagaccaagga 7154 Right cttttggacagttactccgtacc 7155 Left tggttataaatttcaccagcgttgt 7156 Right acagaacaattccaaatgcatataca 7157 Left ttgtagtacagaagttccactgatg 7158 Right ccgtaccttactgagatctggag 7159 Left gatcatcgaaatcaatttgggcaac 7160 Right aatagaggcgagaatttggggaaag 7161 Left tgacttgattagagaccaaggattt 7162 Right gagaatttggggaaagagtggtc 7163 Left aattatgaccaacttgatttgctgt 7164 Right ggtccctgttgttgatgtttgaa 7165 Left cttgatttgctgtttgtctccaag 7166 Right ggtatcctcgtcccaccataaaa 7167 Left cctcattacctggctcactaactaa 7168 Right aatagaggcgagaatttgggga 7169 Left caaaatccattccaattccacagc 7170 Right gcatatagactaaaatcctctgtttgg 7171 Left cgttgtagtacagaagttccactg 7172 Right tgttgttgatgtttgaataaggtaac 7173 Left gtctccttcaaaatccattccaatt 7174 Right ggcttttggacagttactccg 7175 Left cgtctccttcaaaatccattcca 7176 Right agcatatagactaaaatcctctgtt 7177 Left caacttgatttgctgtttgtctcc 7178 Right aggtatcctcgtcccaccata 7179 Left tgcatataaacacaatagaacctgt 7180 Right gcgagaatttggggaaagagtg

TABLE 14 KRAS cDNA Capture Primer List for NGS Panel Seq. ID Primer Sequence KRAS Region1 75-125 bases 7181 Left GCCTGCTGAAAATGACTGAATATAA 7182 Right ATTCGTCCACAAAATGATTCTGAAT 7183 Left GAGAGAGGCCTGCTGAAAATG 7184 Right ATTGTTGGATCATATTCGTCCACAA 7185 Left GGCCTGCTGAAAATGACTGAA 7186 Right TCTATTGTTGGATCATATTCGTCCA 7187 Left GGAGAGAGGCCTGCTGAAA 7188 Right TGGATCATATTCGTCCACAAAATGA 7189 Left GAGAGGCCTGCTGAAAATGACT 7190 Right ATCATATTCGTCCACAAAATGATTC 7191 Left CTCAGCGGCTCCCAGG 7192 Right AAATGATTCTGAATTAGCTGTAT 7193 Left GTGCGGGAGAGAGGCC 7194 Right AAATGATTCTGAATTAGCTG 7195 Left GGGAGAGAGGCCTGCTG 7196 Right TGTTGGATCATATTCGT KRAS Region1 126-175 bases 7197 Left GCCTGCTGAAAATGACTGAATATAAA 7198 Right GTTTCTCCATCAATTACTACTTGCT 7199 Left GAGAGAGGCCTGCTGAAAATG 7200 Right TATTGTTGGATCATATTCGTCCACA 7201 Left GAGAGGCCTGCTGAAAATGACTGAA 7202 Right TCCTCTATTGTTGGATCATATTCGT 7203 Left GGAGAGAGGCCTGCTGAAA 7204 Right TCCATCAATTACTACTTGCTTCCTG 7205 Left GAGAGAGGCCTGCTGAAAATGACT 7206 Right TATTGTTGGATCATATTCGTCCACAA AA 7207 Left CTCAGCGGCTCCCAGG 7208 Right TGGATCATATTCGTCCACAAAATGA 7209 Left GTGCGGGAGAGAGGCC 7210 Right TCAATTACTACTTGCTTCCTGTAGG 7211 Left CCAGGTGCGGGAGAGAG 7212 Right ATCATATTCGTCCACAAAATGATTC 7213 Left GCACTGAAGGCGGCG 7214 Right ATTCGTCCACAAAATGATTCTGAAT 7215 Left GGGAGAGAGGCCTGCTG 7216 Right ATCCTCTATTGTTGGATCATATT 7217 Left GGACTGGGAGCGAGCG 7218 Right AAATGATTCTGAATTAGCTGTAT 7219 Left GCTCCCAGGTGCGGGA 7220 Right AGGAATCCTCTATTGTTGGA 7221 Left AGCGGCTCCCAGGTGC 7222 Right ATCCTCTATTGTTGGATCAT 7223 Left CTGAAAATGACTGAATAT 7224 Right GAATATCCAAGAGACAGGTTTCT 7225 Left CCAGAGGCTCAGCGG 7226 Right AAATGATTCTGAATTAGCTG 7227 Left TCAGCGGCTCCC 7228 Right AGGAATCCTCTATTGTT KRAS Region1 176-225 bases 7229 Left ATTTCGGACTGGGAGCGAG 7230 Right ATTGTTGGATCATATTCGTCCACAA 7231 Left CTCGGCCAGTACTCCCG 7232 Right TGGATCATATTCGTCCACAAAATGA 7233 Left CAGGTGCGGGAGAGAGG 7234 Right TCGAGAATATCCAAGAGACAGGTTT 7235 Left CTCAGCGGCTCCCAGG 7236 Right TCCATCAATTACTACTTGCTTCCTG 7237 Left GCCAGTACTCCCGGCC 7238 Right ATTCGTCCACAAAATGATTCTGAAT 7239 Left TGCGGGAGAGAGGCCT 7240 Right TGTGTCGAGAATATCCAAGAGACAG 7241 Left GCACTGAAGGCGGCG 7242 Right TCAATTACTACTTGCTTCCTGTAGG 7243 Left GCTCCCAGGTGCGGGA 7244 Right GTTTCTCCATCAATTACTACTTGCT 7245 Left CATTTCGGACTGGGAGC 7246 Right TCTATTGTTGGATCATATTCGTCCA 7247 Left AGCGGCTCCCAGGTGC 7248 Right GCTGTGTCGAGAATATCCAAGAG 7249 Left TCCCAGGTGCGGGAGAG 7250 Right CTCTTGACCTGCTGTGTCGA 7251 Left CCAGAGGCTCAGCGG 7252 Right CCTGCTGTGTCGAGAATATCCAA 7253 Left GCTCGGCCAGTACTC 7254 Right TCATATTCGTCCACAAAATGATTCT 7255 Left CCCCGCCATTTCG 7256 Right TCCTCTATTGTTGGATCATATTCGT 7257 Left TCAGCGGCTCCC 7258 Right TTGACCTGCTGTGTCGAGAATATC 7259 Left GGCACTGAAGGCG 7260 Right ATCCTCTATTGTTGGATCATATT 7261 Left CATTTCGGACTGGG 7262 Right AGGAATCCTCTATTGTTGGA 7229 Left CCAGAGGCTCAG 7230 Right GTTTCTCCATCAATTACTACTT KRAS Region2 75-125 bases 7231 Left CTTGGATATTCTCGACACAGCAG 7232 Right TTATGGCAAATACACAAAGAAAGCC 7233 Left AACCTGTCTCTTGGATATTCTCGAC 7234 Right AAATACACAAAGAAAGCCCTCCC 7235 Left GGAAGCAAGTAGTAATTGATGGAGA 7236 Right AATACACAAAGAAAGCCCTCCCCAG 7237 Left AGAAACCTGTCTCTTGGATATTCTC 7238 Right TCCCCAGTCCTCATGTACTG 7239 Left TGTCTCTTGGATATTCTCGACACAG 7240 Right AAAGAAAGCCCTCCCCAGTCC 7241 Left AGCAAGTAGTAATTGATGGAGAAAC 7242 Right TCCCCAGTCCTCATGTA 7243 Left CTACAGGAAGCAAGTAGTAATTGA 7244 Right TCCCCAGTCCTCAT 7245 Left AGAAACCTGTCTCTTGGATATT 7246 Right GATTTAGTATTATTTATGGC 7247 Left AGAAACCTGTCTCTTGGA 7248 Right GGCAAATACACAAAGAAA 7249 Left AGAAACCTGTCTCTT 7250 Right TTTATGGCAAATACACAAAG 7251 Left CAAGTAGTAATTGATGG 7252 Right ATGGCAAATACACA KRAS Region2 126-175 bases 7253 Left TTTGTGGACGAATATGATCCAACAA 7254 Right TTATGGCAAATACACAAAGAAAGCC 7255 Left TGGACGAATATGATCCAACAATAGAG 7256 Right AAATACACAAAGAAAGCCCTCCC 7257 Left CTTGGATATTCTCGACACAGCAG 7258 Right AGGTACATCTTCAGAGTCCTTAAC 7259 Left TTCAGAATCATTTTGTGGACGAATA 7260 Right AATACACAAAGAAAGCCCTCCCCAG 7261 Left CCTTGACGATACAGCTAATTCAGAA 7262 Right TCCCCAGTCCTCATGTACTG 7263 Left CATTTTGTGGACGAATATGATCCAA 7264 Right GAAAGCCCTCCCCAGTCC 7265 Left TGTCTCTTGGATATTCTCGACACAG 7266 Right GTCCTTAACTCTTTTAATTTGTTC 7267 Left AACCTGTCTCTTGGATATTCTCGA 7268 Right TAATTTGTTCTCTATAATGGTGAA 7269 Left GCAAGTAGTAATTGATGGAGAAAC 7270 Right TTTATGGCAAATACACAAAGAAA 7271 Left TGACGATACAGCTAATTCAGAATCA 7272 Right TCCCCAGTCCTCATGTA 7273 Left CAGGAAGCAAGTAGTAATTGATGGA 7274 Right GATTTAGTATTATTTATGGC 7275 Left AGAAACCTGTCTCTTGGATATTCT 7276 Right AATTTGTTCTCTATAATGGT 7277 Left CTACAGGAAGCAAGTAGTAATTG 7278 Right TTTATGGCAAATACACAAAG 7279 Left AGAAACCTGTCTCTTGGATAT 7280 Right GTCCTTAACTCTTTTAATTTG 7281 Left ACAGCTAATTCAGAATCATTTTGTGG 7282 Right TCCCCAGTCCTCAT 7283 Left AGAAACCTGTCTCTTGG 7284 Right ACTCTTTTAATTTGTTCTCT 7285 Left CTACAGGAAGCAAGTAGTAA 7286 Right TATAATGGTGAATATCTTC 7287 Left TCCAACAATAGAGGATTCC 7288 Right TTTATGGCAAATACACA 7289 Left AGAAACCTGTCTCT 7290 Right GTCCTTAACTCTTTTAAT KRAS Region2 176-225 bases 7291 Left CCTTGACGATACAGCTAATTCAGAA 7292 Right TTATGGCAAATACACAAAGAAAGCC 7293 Left TTCAGAATCATTTTGTGGACGAATA 7294 Right GCAAATACACAAAGAAAGCCCTC 7295 Left CAGGAAGCAAGTAGTAATTGATGGA 7296 Right CCATAGGTACATCTTCAGAGTC 7297 Left TGACGATACAGCTAATTCAGAATCA 7298 Right TTTATGGCAAATACACAAAGAAA 7299 Left GGACGAATATGATCCAACAATAGAGG 7300 Right TTTAATTTGTTCTCTATAATGGTG 7301 Left TACAGGAAGCAAGTAGTAA 7302 Right CCATAGGTACATCTTCAGAGTCCTT 7303 Left GCTAATTCAGAATCATTTTGTGGACG 7304 Right TTTATGGCAAATACACAAAG 7305 Left GCAAGTAGTAATTGATGGAGAA 7306 Right GTACATCTTCAGAGTCCTTAAC 7307 Left GTGGACGAATATGATCCAACAATAG 7308 Right AACTCTTTTAATTTGTTCTC 7309 Left CTACAGGAAGCAAGTAGTAATTGA 7310 Right CCATAGGTACATCTTCAGA 7311 Left TCATTTTGTGGACGAATATGATCCA 7312 Right GATTTAGTATTATTTATGGC 7313 Left ACAGCTAATTCAGAATCATTTTGTGG 7314 Right TTTATGGCAAATACACA 7315 Left TCCAACAATAGAGGATTCC 7316 Right GTCCTTAACTCTTTTAATTTGTT 7317 Left GAATCATTTTGTGGACGAATATGA 7318 Right ATAATGGTGAATATCTTC 7319 Left AGAAACCTGTCTCTTG 7320 Right CTAGAAGGCAAATCACATTTATTT 7321 Left TACAGGAAGCAAGTAG 7322 Right GTCCTTAACTCTTTTAATTT 7323 Left GAATATGATCCAACAA 7324 Right GTCCTTAACTCTTTTAA 7325 Left CTACAGGAAGCAAG 7326 Right CCATAGGTACATCTTC KRAS Region3 176-225 bases 7327 Left gactgaatataaacttgtggtagt 7328 Right tccccagtcctcatgtactg 7329 Left gactgaatataaacttgtggt 7330 Right tccccagtcctcatgta 7331 Left tgactgaatataaacttgt 7332 Right ccccagtcctcat KRAS Region3 226-275 bases 7333 Left atgactgaatataaacttgtggtag 7334 Right ttatggcaaatacacaaagaaagcc 7335 Left GCCTGCTGAAAatgactgaatataaa 7336 Right atacacaaagaaagccctcccc 7337 Left GGAGAGAGGCCTGCTGAAAat 7338 Right gcaaatacacaaagaaagccctc 7339 Left CCTGCTGAAAatgactgaatataaactt 7340 Right tccccagtcctcatgtactg 7341 Left GAGAGGCCTGCTGAAAatgact 7342 Right atacacaaagaaagccctccccagt 7343 Left GGGAGAGAGGCCTGCTG 7344 Right caaagaaagccctccccagtcct 7345 Left GGCCTGCTGAAAatgactgaata 7346 Right ccccagtcctcatgta 7347 Left TGCGGGAGAGAGGCCT 7348 Right tatggcaaatacacaaagaaa 7349 Left Aatgactgaatataaacttgtgg 7350 Right tccccagtcctcat 7351 Left GGTGCGGGAGAGAGG 7352 Right ggcaaatacacaaag KRAS Region3 276-325 bases 7353 Left CTCAGCGGCTCCCAGG 7354 Right ttatggcaaatacacaaagaaagcc 7355 Left tgactgaatataaacttgtggtagt 7356 Right gtccttaactcttttaatttgttc 7357 Left TCCCAGGTGCGGGAGA 7358 Right aaatacacaaagaaagccctccc 7359 Left GGCCTGCTGAAAatgactgaat 7360 Right tttaatttgttctctataatggtg 7361 Left ATTTCGGACTGGGAGCGAG 7362 Right tccccagtcctcatgtactg 7363 Left GCACTGAAGGCGGCG 7364 Right aatacacaaagaaagccctccccag 7365 Left TGCGGGAGAGAGGCCT 7366 Right tttatggcaaatacacaaagaaa 7367 Left GGCCTGCTGAAAatgactgaatata 7368 Right actcttttaatttgttctct 7369 Left AGCGGCTCCCAGGTGC 7370 Right cacaaagaaagccctccccagtcc 7371 Left CCTGCTGAAAatgactgaatataaact 7372 Right gatttagtattatttatggc 7373 Left tgactgaatataaacttgtggt 7374 Right acatcttcagagtccttaa 7375 Left GAGAGAGGCCTGCTGAAAatg 7376 Right tataatggtgaatatcttc 7377 Left CATTTCGGACTGGGAGC 7378 Right tccccagtcctcatgta 7379 Left CCAGAGGCTCAGCGG 7380 Right tttatggcaaatacacaaag 7381 Left CCAGGTGCGGGAGAGAG 7382 Right tttatggcaaatacaca 7383 Left CTGAAAatgactgaatataaacttgt 7384 Right gtccttaactcttttaa 7385 Left GGCCTGCTGAAAatgact 7386 Right ccttaactcttttaatttg 7387 Left GGCACTGAAGGCG 7388 Right tccccagtcctcat KRAS Region3 326-375 bases 7389 Left ATTTCGGACTGGGAGCGAG 7390 Right ttatggcaaatacacaaagaaagcc 7391 Left CTCAGCGGCTCCCAGG 7392 Right ccataggtacatcttcagagtcctt 7393 Left CTCGGCCAGTACTCCCG 7394 Right aaatacacaaagaaagccctccc 7395 Left GTGCGGGAGAGAGGCC 7396 Right taggtacatcttcagagtccttaac 7397 Left GCCAGTACTCCCGGCC 7398 Right aatacacaaagaaagccctccccag 7399 Left CCAGGTGCGGGAGAGAG 7400 Right ccataggtacatcttcagagtc 7401 Left GCTCCCAGGTGCGGGA 7402 Right gtccttaactcttttaatttgac 7403 Left GCACTGAAGGCGGCG 7404 Right taatttgttctctataatggtgaa 7405 Left CATTTCGGACTGGGAGC 7406 Right aaagaaagccctccccagtcc 7407 Left AGCGGCTCCCAGGTGC 7408 Right gtccttaactcttttaatttg 7409 Left GCTCGGCCAGTACTC 7410 Right tatggcaaatacacaaagaaa 7411 Left CCAGAGGCTCAGCGG 7412 Right actcttttaatttgttctct 7413 Left CCCCGCCATTTCG 7414 Right tttatggcaaatacacaaag 7415 Left TCAGCGGCTCCC 7416 Right ccataggtacatcttcaga 7417 Left GGCACTGAAGGCG 7418 Right aatttgttctctataatggt KRAS Region3 376-425 bases 7419 Left CCAGGTGCGGGAGAGAG 7420 Right actgttctagaaggcaaatcacatt 7421 Left CTCAGCGGCTCCCAGG 7422 Right tctactgttctagaaggcaaatcac 7423 Left TTTCGGACTGGGAGCGAG 7424 Right aggtacatcttcagagtccttaac 7425 Left CTCGGCCAGTACTCCCG 7426 Right tttatggcaaatacacaaagaaagcc 7427 Left GTGCGGGAGAGAGGCC 7428 Right gttttgtgtctactgactagaagg 7429 Left GCACTGAAGGCGGCG 7430 Right ccataggtacatcttcagagtcctt 7431 Left TCCCAGGTGCGGGAGA 7432 Right tgttctagaaggcaaatcacatttat 7433 Left AGCGGCTCCCAGGTGC 7434 Right gcctgttttgtgtctactgttc 7435 Left GCCAGTACTCCCGGCC 7436 Right taatttgttctctataatggtgaa 7437 Left CATTTCGGACTGGGAGC 7438 Right gtccttaactcttttaatttgttc 7439 Left CCAGAGGCTCAGCGG 7440 Right ccataggtacatcttcagagtc 7441 Left GCTCGGCCAGTACTC 7442 Right ttatggcaaatacacaaagaaagccctc 7443 Left AAGGTGGCGGCG 7444 Right aaatacacaaagaaagccctcccc 7445 Left GGCGGCGAAGGT 7446 Right atacacaaagaaagccctccccagt 7447 Left GCGAAGGTGGCG 7448 Right caaagaaagccctccccagtcct 7449 Left TCAGCGGCTCCC 7450 Right tctactgttctagaaggcaaat 7419 Left CGGAGGCAGCAG 7420 Right tttatggcaaatacacaaagaaa 7421 Left CCCCGCCATTTCG 7422 Right gtccttaactcttttaatttg 7423 Left GGCACTGAAGGCG 7424 Right ccataggtacatcttcaga 7425 Left CATTTCGGACTGGG 7426 Right actcttttaatttgttctct KRAS Region3 426-475 bases 7427 Left ATTTCGGACTGGGAGCGAG 7428 Right actgttctagaaggcaaatcacatt 7429 Left GCTCGGCCAGTACTCCC 7430 Right ccataggtacatcttcagagtcctt 7431 Left GTGCGGGAGAGAGGCC 7432 Right gtcttgtctttgctgatgtttcaat 7433 Left GCCAGTACTCCCGGCC 7434 Right taggtacatcttcagagtccttaac 7435 Left CTCAGCGGCTCCCAGG 7436 Right agcctgttttgtgtctactgttc 7437 Left GCACTGAAGGCGGCG 7438 Right tctactgttctagaaggcaaatcac 7439 Left CCAGGTGCGGGAGAGAG 7440 Right cttcttgctaagtcctgagcct 7441 Left GCTCCCAGGTGCGGGA 7442 Right tgctgatgtttcaataaaaggaattcc 7443 Left CCAGAGGCTCAGCGG 7444 Right gttttgtgtctactgttctagaagg 7445 Left AGCGGCTCCCAGGTGC 7446 Right ttgctgatgtttcaataaaaggaat 7447 Left CATTTCGGACTGGGAGC 7448 Right tgttctagaaggcaaatcacatttat 7449 Left GGCACTGAAGGCG 7450 Right ttcttgctaagtcctgagcctgttt 7451 Left GCTCGGCCAGTACT 7452 Right ccataggtacatcttcagagtc 7453 Left AAGGTGGCGGCG 7454 Right gtccttaactcttttaatttgttc 7455 Left GGCGGCGAAGGT 7456 Right taatttgttctctataatggtgaa 7457 Left TCAGCGGCTCCC 7458 Right gaattccataacttcttgctaag 7459 Left CATTTCGGACTGGG 7460 Right tctactgttctagaaggcaaat 7461 Left CCCCGCCATTTCG 1462 Right ccataggtacatcttcaga 7463 Left GCGAAGGTGGCG 7464 Right gtccttaactcttttaatttg 7465 Left CCAGAGGCTCAG 7466 Right agcctgttttgtgtctactg KRAS Region3 476-525 bases 7467 Left GCTCGGCCAGTACTCCC 7468 Right actgttctagaaggcaaatcacatt 7469 Left CATTTCGGACTGGGAGCGA 7470 Right gttttgtgtctactgttctagaagg 7471 Left CAGGTGCGGGAGAGAGG 7472 Right atgtatagaaggcatcatcaacacc 7473 Left CTCAGCGGCTCCCAGG 7474 Right gtcttgtctttgctgatgtttcaat 7475 Left TCCCAGGTGCGGGAGA 7476 Right tatagaaggcatcatcaacaccctg 7477 Left TGCGGGAGAGAGGCCT 7478 Right atcatcaacaccctgtcttgtcttt 7479 Left GCCAGTACTCCCGGCC 7480 Right agcctgttttgtgtctactgttc 7481 Left GCACTGAAGGCGGCG 7482 Right ctgtcttgtctttgctgatgtttc 7483 Left GGACTGGGAGCGAGCG 7484 Right ttcttgctaagtcctgagcctgttt 7485 Left AGCGGCTCCCAGGTGC 7486 Right catcatcaacaccctgtcttgtc 7487 Left CCAGAGGCTCAGCGG 7488 Right tgctgatgtttcaataaaaggaattcc 7489 Left GCTCGGCCAGTACT 7490 Right tctactgttctagaaggcaaatcac 7491 Left CATTTCGGACTGGGAG 7492 Right cttcttgctaagtcctgagcct 7493 Left GGCGGCGAAGGT 7494 Right ccataggtacatcttcagagtcctt 7495 Left AAGGTGGCGGCG 7496 Right tgttctagaaggcaaatcacatttat 7497 Left GGCACTGAAGGCG 7498 Right tcttgtctttgctgatgtttcaataaa 7499 Left TCAGCGGCTCCC 7500 Right ttgctgatgtttcaataaaaggaat 7501 Left CGGAGGCAGCAG 7502 Right taggtacatcttcagagtccttaac 7503 Left CCCCGCCATTTCG 7504 Right tctactgttctagaaggcaaat 7505 Left GCGAAGGTGGCG 7506 Right gttttgtgtctactgttctaga KRAS Region3 526-575 bases 7507 Left ATTTCGGACTGGGAGCGAG 7508 Right atgtatagaaggcatcatcaacacc 7509 Left GCTCGGCCAGTACTCCC 7510 Right gtcttgtctttgctgatgtttcaat 7511 Left GCCAGTACTCCCGGCC 7512 Right ctgtcttgtctttgctgatgtttc 7513 Left GTGCGGGAGAGAGGCC 7514 Right ttttaccatctttgctcatcttttc 7515 Left GCACTGAAGGCGGCG 7516 Right tatagaaggcatcatcaacaccctg 7517 Left CATTTCGGACTGGGAGC 7518 Right atcatcaacaccctgtcttgtcttt 7519 Left CTCAGCGGCTCCCAGG 7520 Right ttttaccatctttgctcatctt 7521 Left GCTCGGCCAGTACT 7522 Right tgctgatgtttcaataaaaggaattcc 7523 Left AGCGGCTCCCAGGTGC 7524 Right tgctcatcttttctttatgttt 7525 Left GGCGGCGAAGGT 7526 Right gttttgtgtctactgttctagaagg 7527 Left CCCCGCCATTTCG 7528 Right tcttgtctttgctgatgtttcaataaa 7529 Left AAGGTGGCGGCG 7530 Right ttgctgatgtttcaataaaaggaat 7531 Left GGCACTGAAGGCG 7532 Right catcatcaacaccctgtcttgtc 7533 Left GCTCCCAGGTGCGGGA 7534 Right ctttatgttttcgaatttctc 7535 Left CGGAGGCAGCAG 7536 Right actgttctagaaggcaaatcacatt 7537 Left GCGAAGGTGGCG 7538 Right agcctgttttgtgtctactgttc 7539 Left CCAGGTGCGGGAGAGAG 7540 Right tttaccatctttgctcat 7541 Left CCAGAGGCTCAGCGG 7542 Right gaatttctcgaactaatgtata 7543 Left CATTTCGGACTGGG 7544 Right cctgtcttgtctttgctgatgt 7545 Left TCAGCGGCTCCC 7546 Right ttgctcatcttttctttatg KRAS Region3 576-625 bases 7547 Left GCTCGGCCAGTACTCCC 7548 Right atgtatagaaggcatcatcaacacc 7549 Left CATTTCGGACTGGGAGCGA 7550 Right ttttaccatctttgctcatcttttc 7551 Left GCCAGTACTCCCGGCC 7552 Right tatagaaggcatcatcaacaccctg 7553 Left GGACTGGGAGCGAGCG 7554 Right ttttaccatctttgctcatctt 7555 Left GCACTGAAGGCGGCGG 7556 Right tgctcatcttttctttatgttt 7557 Left GCTCGGCCAGTACT 7558 Right atcatcaacaccctgtcttgtcttt 7559 Left AAGGTGGCGGCG 7560 Right gtcttgtctttgctgatgtttcaat 7561 Left GGCGGCGAAGGT 7562 Right ctgtcttgtctttgctgatgtttc 7563 Left GCGAAGGTGGCG 7564 Right tgctgatgtttcaataaaaggaattcc 7565 Left CAGGTGCGGGAGAGAGG 7566 Right ttacataattacacactttg 7567 Left CCCCGCCATTTCG 7568 Right taatgtatagaaggcatcatcaac 7569 Left CTCAGCGGCTCCCAGG 7570 Right cacactttgtctttgac 7571 Left CGGAGGCAGCAG 7572 Right cctgtcttgtctttgctgatgt 7573 Left CATTTCGGACTGGGAG 7574 Right gaatttctcgaactaatgtata 7575 Left GGCACTGAAGGCGG 7576 Right ttttaccatctttgctcat 7577 Left CTCCCAGGTGCGGGA 7578 Right gtctttgacttcttt 7579 Left CATTTCGGACTGG 7580 Right ctttatgttttcgaatttctc KRAS Region3 626-675 bases 7581 Left GCTCGGCCAGTACTCCC 7582 Right ttttaccatctttgctcatcttttc 7583 Left GCCAGTACTCCCGGCC 7584 Right ttttaccatctttgctcatctt 7585 Left ATTTCGGACTGGGAGCGAG 7586 Right ttacataattacacactttg 7587 Left CGGAGGCAGCAG 7588 Right atgtatagaaggcatcatcaacacc 7589 Left GCTCGGCCAGTACT 7590 Right tgctcatcttttctttatgttt 7591 Left CATTTCGGACTGGGAGC 7592 Right cacactttgtctttgac 7593 Left GGCGGCGAAGGT 7594 Right gaatttctcgaactaatgtata 7595 Left AAGGTGGCGGCG 7596 Right ctttatgttttcgaatttctc 7597 Left CCCCGCCATTTCG 7598 Right ttttaccatctttgctcat 7599 Left GCGAAGGTGGCG 7600 Right ttgctcatcttttctttatg 7601 Left CATTTCGGACTGGG 7602 Right gtctttgacttcttt KRAS Region3 676-725 bases 7603 Left GCTCGGCCAGTACTCCC 7604 Right ttacataattacacactttg 7605 Left CGGAGGCAGCAG 7606 Right ttttaccatctttgctcatctttt 7607 Left GCTCGGCCAGTACT 7608 Right cacactttgtctttgac 7609 Left AAGGTGGCGGCG 7610 Right gtctttgacttcttt

TABLE 15 IRID Hybridization-based capture probes for NGS panel ID Probe Sequence 7611 CGGTCCAGAGCCAAGCGGCGGCAGAGCGAGGGGCATCAGCTACCGCCAAGTCCAGAG CCATTTCCATCCTGCAGAAGAAGCCCCGCCACCAG 7612 AGAAATGGATACAGGTCAAGTCTAAGTCGAATCCATCCTCTTGATATCTCCTTTTGTTTC TGCTAACGATCTCTTTGATGATGGCTGTCATGTCTGGGAGCCTGTGGCTGAAGAAAAAG GAGGAGAGAGATGGCAGAAGCTGC 7613 AAGTTGACAGGGTACCAGGAGATGATGTAAGGGACAAGCAGCCACACCCCATTCTTGA GGGGCTGAGGTGGAAG 7614 GATTGGAGACTTCGGGATGGCCCGAGACATCTACAGGTGAGTAAAGACTGCCTCACCC CTCCGGGCCTGTCTCTTCCACCTCAGCCCCTCAAGAATGGGG 7615 CTTGGCCACTCTTCCAGGGCCTGGACAGGTCAAGAGGCAGTTTCTGGCAGCAATGTCTC TGGGAAGAAAGGAAATGCATTTCCTAATTTTATCCCTAGGAAGATGAGTGTACAACGG 7616 CACTTTGACTCACCGGTGGATGAAGTGGTTTTCCTCCAAATACTGACAGCCACAGGCAA TGTCCCGAGCCACGTGCAGAAGGTCCAGCATGGCCAGGGAGGAGGGCTGGCTCTGTGG GGAGACAGAAGCGGGC 7617 CCCTGGGTTCCATCGAGGAACTTGCTACCCAGGCTGCCCACTCTTGCTCCTTCCATCCTT GCTCCTGTCCTTGGC 7618 GTTCATCCTGCTGGAGCTCATGGCGGGGGGAGACCTCAAGTCCTTCCTCCGAGAGACCC GCCCTCGCCCGGTGAGTGAGAACCAGTCTTTGCTGCAGTTGTTGTGCC 7619 CCGGGGCAGGGATTGCAGGCTCACCCCAATGCAGCGAACAATGTTCTGGTGGTTGAAT TTGCTGCAGAGCAGAGAGGGATGTAACCAAAATTAACTGAGCTGAGTCTGGGCAAATC TTAAACTGGGAGGAACA 7620 GGGAGGGCTCTGCCGGCCTTTTGTGGCTAGAGGAGTCTGCGGTGCTGTGATAACATTCA GCCCCTACACTGCACCCCTCTCCTCCCAGGACGGC 7621 CCCAACTACTGCTTTGCTGGCAAGACCTCCTCCATCAGTGACCTGAAGGAGGTGCCGCG GAAAAACATCACCCTCATTCGGTGAGCGCCCTGCTGC 7622 GGTTGTAGTCGGTCATGATGGTCGAGGTGCGGAGCTTGCTCAGCTTGTACTCAGGGCTC TGCAGCTCCATCTGCATGGCTTGCAGCTCCTGGTGCTTCCGGCGGTACACTGCAGGTGG GTG 7623 TTTTAAATACCTGTTAAGTTTGTATGCAACATTTCTAAAGTTACCTACTTGTTAATTAAA AATTCAAGAGTTTTTTTTTCTTATTCTGAGGTTATCTTTTTACCACAG 7624 TTAGCCATTGGTCAAGATCTTCACAAAAGGGTTTGATAAGTTCTAGCTGTGGTGGGTTA TGGTCTTCAAAAGGATATTGTGCAACT 7625 AAGTGAAGATGACAATCATGTTGCAGCAATTCACTGTAAAGCTGGAAAGGGACGAACT GGTGTAATGATATGTGCATATTTATTACATCGGGGCAA 7626 GAAGAGGAAAGGAAAAACATCAAAAAATAACTTACCTTTTTGTCTCTGGTCCTTACTTC CCCATAGAAATCTAGGGCCTCTTGTGCCTTTAAAAATT 7627 CAGTTACCATAGCAATTTAGTGAAATAACTATAATGGAACATTTTTTTTCAATTTGGCTT CTCTTTTTTTTCTGTCCACCAGGGAGTAACTA 7628 ACATCATCTTGTGAAACAACAGTGCCACTGGTCTATAATCCAGATGATTCTTTAACAGG TAGCTATAATAATACACATAGCGCCTCTGACTGGGAAT 7629 TTTGAAACTATTCCAATGTTCAGTGGCGGAACTTGCAGTAAGTGCTTGAAATTCTCATC CTTCCATGTATTGGAACAGTTTTCTTAACCATATCTAGAAGTTTACATAAAAATTTAGA AAGAAATTT 7630 ATATTTCGTGTATATTGCTGATATTAATCATTAAAATCGTTTTTGACAGTTTGACAGTTA AAGGCATTTCCTGTGAAATAATAC 7631 CGTGTGGGTCCTGAATTGGAGGAATATATCTTCACCTTTAGCTGGCAGACCACAAACTG AGGATCTGCATGGTTAAATACATACCAGTATT 7632 GACGGGAAGACAAGTTCATGTACTTTGAGTTCCCTCAGCCGTTACCTGTGTGTGGTGAT ATCAAAGTAGAGTTCT 7633 TTATAGTTCCTTACATGTCATAAAATAAAATATAGCTTTTAATCTGTCCTTATTTTGGAT ATTTCTCCCAATGAAAGTAAAGTACAAACCTTTTTTAGCATCTTGTTCTGTTTGTGGAA 7634 TAACATAGGTGACAGATTTTCTTTTTTAAAAAAATAAAACATCATTAATTAAATATGTC ATTTCATTTCTTTTTCTTTTCTTTTTTTTTTTTTTTAGGACAAAATGTTTCACTTTTGGGTA 7635 ATCACATAGACTTCCATTTTCTACTTTTTCTGAGGTTTCCTCTGGTCCTGGTATGAAGAA TGTATTTA 7636 TATGTGATCAAGAAATCGATAGCATTTGCAGTATAGAGCGTGCAGATAATGACAAGGA ATATCTAGTACTTACTTTAACAAAAAATGATCTTGACAAAGCAAATAAAGACAA 7637 TACAAGTCAACAACCCCCACAAAATGTTTAATTTAACTGACCTTAAAATTTGGAGAAAA GTATCGGTTGGCTT 7638 AGAGAAACCTTTATCTGTATCAAAGAATGGTCCTGCACCAGTAATATGCATATTAAAAC AAGATTTACCTCTATTGTTGGATCA 7639 CTGGTGGCGTAGGCAAGAGTGCCTTGACGATACAGCTAATTCAGAATCATTTTGTGGAC GAATATG 7640 GCTCCAACTACCACAAGTTTATATTCAGTCATTTTCAGCAGGCCTTATAATAAAAATAA TGAAAATGTGACTATATTAGAACATGTCACACATAAGGTTAATACA 7641 GCCCATGCCGTGGCTGCTGGTCCCCCTGCTGGGCCATGTCTGGCACTGCTTTCCAGCAT GGTGAGGGCTGAGGTGACCCTTGTCTCTGTGTTCTTGTCCCCCCCAGCTTGTGGAGCCT CTTACACC 7642 CAGAGGCCTGTGCCAGGGACCTTACCTTATACACCGTGCCGAACGCACCGGAGCCCAG CACTTTGATCTTTTTGAATTCAGTTTCCTTCAAGATCCTCAAGAGAGCTTGGTTGGGAGC TTCTCCACTGG 7643 CCTTCGGGGTGCATCGCTGGTAACATCCACCCAGATCACTGGGCAGCATGTGGCACCAT CTCACAATTGCCAGTTAACGTCTTCCT 7644 TCTCTTAATTCCTTGATAGCGACGGGAATTTTAACTTTCTCACCTTCTGGGATCCAGAGT CCCTATGACAGAGAGAGAAG 7645 AAGCAACATCTCCGAAAGCCAACAAGGAAATCCTCGATGTGAGTTTCTGCTTTGCTGTG TGGGGGTCCATGGCTCTGAACCTCAGGCCCACCTTTTCTCATGTCTGGCAGC 7646 ATTTTGAAACTCAAGATCGCATTCATGCGTCTTCACCTGGAAGGGGTCCATGTGCCCCT CCTTCTGGCCACCATGCGAAG 7647 GAGGTGAGGCAGATGCCCAGCAGGCGGCACACGTGGGGGTTGTCCACGCTGGCCATCA CGTAGGCTTCCTGGAGGGAGGGAGAGGCACGTCAGTGTGGC 7648 CCACCGTGCAGCTCATCACGCAGCTCATGCCCTTCGGCTGCCTCCTGGACTATGTCCGG GAACACAAAGACAATATTGGCTCCCAGTACCTGCTCAACTGGTGTGTGCAGATCGCAA AGGTAATCAGGGAAGGGA 7649 CGTGGAGAGGCTCAGAGCCTGGCATGAACATGACCCTGAATTCGGATGCAGAGCTTCT TCCCATGATGATCTGTCCCTCACAGCAGGGT 7650 GCGGTGTTTTCACCAGTACGTTCCTGGCTGCCAGGTCGCGGTGCACCAAGCGACGGTCC TCCAAGTAGTTCATGCCCTGAAACAGAGAAGACCCTGC 7651 CAGCATGTCAAGATCACAGATTTTGGGCTGGCCAAACTGCTGGGTGCGGAAGAGAAAG AATACCATGCAGAAGGAGGCAAAGTAAGGAGGTGGCTTTAGGTCAGCCAGCATTT 7652 GATTATCTGTCTGGCCCCAGACCTGGAGCTTTCTTTCCATGATAGGAGTACTTCTTTGGG TTGACTTCTCTGGTGACAG 7653 CTCATCAAGCTCTAGCTCCTCCAGCTTCTTCTGCAAGGCCTCCAAGTTGGTCCTGTTCCA AAGTGGGGAGCACAAGTCAATACT 7654 GCAGCAGCGAAAGCGCCTTGAGGCCTTTCTTACCCAGAAGCAGAAGGTGGGAGAACTG AAGGATGACGACTTTGAGAAGATCAGTGAGCTGGGGGCTGGCAATGGCGGTGTGGTGT TCAAG 7655 CCCCAGGCTTCTAAGTACCCTGAGAAATAATCCAATTACCTGTTAATCAAGGCAAACTC ACCTTTCTGGCCATGACCAGGCCAGAAGGCTTGTGGGAGACC 7656 AAAATACTATAGTTGAGACCTTCAATGACTTTCTAGTAACTCAGCAGCATCTCAGGGCC AAAAATTTAATCAGTGGAAAAATAGCCTCA 7657 CTACAGTGAAATCTCGATGGAGTGGGTCCCATCAGTTTGAACAGTTGTCTGGATCCATT TTGTGGATGGTAAGAATT 7658 GCTAGACCAAAATCACCTATTTTTACTGTGAGGTCTTCATGAAGAAATATATCTGAGGT GTAGTAAGTAAAGGAAAACAGTAGAT 7659 ATATATACATAAGAGAGAAGGTTTGACTGCCATAAAAAATATCTAATTTATGACAATA AAAACCTTACTTTATTTGGATTTGAT 7660 ATATATAATAGCTTTTCTTCCATCTCTTAGGAAACTCCATGCTTAGAGTTGGAGTTTGAC TGGTTCAGCAGTGTGGTAAAGT 7661 TTATAATTGATACTTAATAAACTCAGTGATTTGCCTTACCAGTCCTGCGTGGGAATAGC TAAATCCTGCTTCTCGGGATACAGACCAATTGGCATGCTCTTCAATCACTGACATATCT GGGAA 7662 TTTCTCCTGCAGCTGTGTGGACCTGGATGACAAGGGCTGCCCCGCCGAGCAGAGAGCC AGGTTGGCCTGGACCC 7663 GCTAGGGACAACACGATTTCCCTTGGAGATATCGATCTGTTAGAAACCTCTCCAGGTTC TTTGGGGGCAGAG 7664 TCATGGAAGCCCTGATCATCAGGTAAAGCCACAGAGAGACACCCTCACCCCAACTCCC CTCTGCCCCCAAAGAAC 7665 GAGGAAATCCAGTTCGTCCTGTTCAGAGCACACTTCAGGCAGCGTCTGGGCAGAGAAG GGGAGGGTGGGGAGGAGGAGGAGGCTGTGA 7666 CAAGTGGCTGTGAAGGTAAGAAGTGGCTCACTCTTGAGCCTGCCCTTGGCTTGCGGACT CTGTAGGCTGCAGTTCTCAGCTC 7667 GCAGGGGGCTTGGGTCGTTGGGCATTCCGGACACCTGGCCTTCATACACCTCCCCAAAG GCGCCATGGCCCAGACCCCTGTGCAAAGGAGAAGACAAGAGG 7668 CCCTCTAGGGTTGTCAATGAAATGAATTCACCAACATAAAATGGTTTTGAAAAATCCTA AAGAGCTCTACCAATGTGAGTGACCATTATCACTCCT 7669 GCTGGGCTTTACACACAGAATCTACCCACTGAATCACAATTTTGTTCTGGCTTCCATGG AGTTTGCCTTCCAGAACATCCTCACATGTA 7670 CCCCCCAACACATGGGCCAGGGCAAATGAGTCACCCGCTATGTGCTCAGTTCCCTCCTC TATGCAATGGACCGACCGTGATCAGATTAGGGTTACCTGAGGATCGAATGAATTGAAA TG 7671 TAATATCACATTGTATAGACATACTGTGTTTTTAAAATCTATTCATCAGTTGAGGAACA GTTTTAGAAAATGAATAGTTGTTAAACCTGAATT 7672 AGAACCTTATTGAAATATGGCCAAAGTAAGTCAATAAACAGCCCTTAGTCCTCTTGAAA AAATTCAGGTTTAACAACTATTCATTTT 7673 TACCTTCCCATTTTGAACCTGGTAACTAATTCTGTAGGACATCATCTTCCAAAAAGCTTT ACTCTTGCTCTGCTAGCCTATCTAGCTAAAGAGCTTCTCTG 7674 GGCCACTTCCCAGCTGGCGCGGACACGGCAGGCTGGAGAGCCATGAGGCAGAGCATAC GCAGCCTGTACCCAGTGGTGCCGAGCCTCTGGCG 7675 CGATGAAGGAGAAGAGGACAGCGGCTGCGATCACCGTGCGGCACAGCTCGTCGCACA GTGGATCTGTGGGTGGGGGTGGTGTGAGGCTTGGCACCG 7676 CATCGTCTCGGTGCTGCTGTCTGCCTTCTGCATCCACTGCTACCACAAGTTTGCCCACAA GCCACCCATCTCCTCAGCTGAGATGAC 7677 GAGACCTGGTTCTCCATGGAGTCCAGCGAGGGCCGGCGGGCACCGGAAGAGGAGTAGC TGACCGGGAAGGCCTGGGCGGGCCTCCGGAAGG 7678 CCGTGGATGCCTTCAAGATCCTGGTGAGGGTCCCTGCGGGGCAGGGAAGATCCCCTGC CCTCCCCAGCTGCCTTCCAGGGAGGGAGGCCAGCTGG 7679 CCAGGAGTGTCTACAGCACTCCTCTGGTTACTGAAAGCTCAGGGATAGGGCCTGGCCTT CTCCTTTACCCCTCCTTCCTAGAGAGTTAGA 7680 AAAAGGGATTCAATTGCCATCCATTTAACTGGAATCCGACCCTAAAGAGAAGATGGAA TAAAGACATTGAAGTTACTC 7681 TTGATCATATCTACACCACGCAAAGTGATGTGTAAGTGTGGGTGTTGCTCTCTTGGGGT GGAGGTTACAGAAACACCCTTATACATGTAGTGGGGCCACGACGCCCGTCTGTGCAGC TTGGCCAG 7682 TCTTATTCCTTTAATACAGAATATGGGTAAAGATGATCCGACAAGTGAGAGACAGGAT CAGGTCAGCGGGCTACCACTGGGCC 7683 CAGGTGGTGTTGGGAAAAGCGCACTGACAATCCAGCTAATCCAGAACCACTTTGTAGA TGAATATGATCCCACCATAGAGGTGAGG 7684 GCTCCAACCACCACCAGTTTGTACTCAGTCATTTCACACCAGCAAGAACCTGTTGGAAA CCAGTAATCAGGGTTAATTGGCGAGCCACATCTACAGTACTTTAAAGCTTTCTATAATCA 7685 AAGAAGAGTACAGTGCCATGAGAGACCAATACATGAGGACAGGCGAAGGCTTCCTCTG TGTATTTGCCAT 7686 GTACTCTTCTTGTCCAGCTGTATCCAGTATGTCCAACAAACAGGTTTCACCATCTATAAC CACTTGTTTTCTGTAAGAATCCTGGGGGTGT 7687 CTTGGCAATAGCATTGCATTCCCTGTGGTTTTTAATAAAAATTGAACTTCCCTCCCTCCC TGCCCCCTTACCCTCCA 7688 GGTTAAATAAGCATCTAACTATTCAAGCCCATTTCTGCCTATCTGGTTTGTCCCTCAAAT TGCTAATATATAATCACAAACAAAAAGTATCCAATATCACCCTACATAAAAGAAAACCC 7689 GGGCTTTCTTTGTGTATTTGCCATAAATAATACTAAATCATTTGAAGATATTCACCATTA TAGGTGGGTTTAAATTGAATATAATAAGCTGACATTAAGGAGTAATTATAGTTTTTATT TTTTGAG 7690 CCTCCCCAGTCCTCATGTACTGGTCCCTCATTGCACTGTACTCCTCTTGACCTGCTGTGT CGAGAATATCCAAGAGACAGGT 7691 TTTTTAAATTGAATTTTTTGTTGTTGAGTTGTATATAACACCTTTTTTGAAGTAAAAGGT GCACTGTAATAATCCAGACTGTGTTTCTCCCTTCTCAGGATTCCTACAGGAAGCAAGTA GTAATTGATGGAGAAA 7692 GCTTGTTGCTTGCCAGCCCAGGACTTGGAGGCTCCAGGGGACCCCCATCGTGGGGCCTG GTGGGCAAAGAGGGCTCCAGCCAACCCCCCAAAT 7693 CTACAAGGAGCGGCCGCAGGATGTGGACCAACGTGAGGCTCCCCTCAACAACTTCTCT GTGGCGCGTAAGTATCCCCTTGGCCTCTCGGGATTCAGATTTGGGGGGTTGGCTGGAGC CC 7694 GCCAATGAAGGTGCCATCATTCTTGAGGAGGAAGTAGCGTGGCCGCCAGGTCTTGATG TACTCCCCTACAGACGTGCGGGTGGTGAGAGCCACGCACACTCTACCCGTCAGACCCTC GCCAGGCAGCCAGG 7695 AAACTAATTTTTGAGACAAGATAATTTTTTATAAATAAATATTTCAGAATTCTAAGGTC AAAATTAGAACAGTAGATGCTTAGTTTA 7696 TTTCATGCCTTTGGCTACTTGAAGACCAAAGCCAATAAGATCTTTTACAGTTGGATTCT GCAGTCAAAGAGAGTTAGAAAAGCAT 7697 ATATCTTGCAAGCAAAAAGTTTGTCCACAGAGACTTGGCTGCAAGAAACTGTATGTAA GTATCAGAATCTCTGTGCCACAATCCAAATTAAGTGACAAGGAGGAATCTG 7698 AAGTAGTAATAACAGTGCTGTTGATATTGAGACAACACCAGCAATCAATCCTGTGAAA TTCTGATCTGGTTGAA 7699 TTGGGTTTTTCCTGTGGCTGAAAAAGAGAAAGCAAATTAAAGGTGCATTTTTGTTACTG TTCATTTTTAGAAGTTA 7700 GGTTTTATGGACTACATATAAGACAGCACACAAGAATCGACGACAATCTTAAACTGTA ATGACTGTGTTCTTAAGGTA 7701 ATAAAACCCATGAGTTCTGGGCACTGGGTCAAAGTCTCCTGGGGCCCATGATAGCCGTC TTTAACAAGCTCTTT 7702 CTTCGGGCACTTACAAGCCTATCCAAATGAGGAGTGTGTACTCTTGCATCGTAGCGAAC TAATTCACTGCCCAGATCTTAAAACAGAGAGAAAGA 7703 GTGTAAGCCCAACTACAGAAATGGTTTCAAATGAATCTGTAGACTACCGAGCTACTTTT CCAGAAGGTATATTTCAGTTTATTGTTCTGAGAAATACCTATACAT 7704 GCAGCAGCGAAAGCACCTTGAGGCCTTTCTTACCCAGAGATAGAAGGTGGGAGAACTA AAGGATGACGACTTTGAGAAGATCAGTGAGCTGGGGGCTGGCAATGGCGGTGTGGTGT CCAAA 7705 TATGTGATCAAGAAATTGATAGCATTTGCAGTATAGAGCGTGCAGATAATGACAAGGA GTATCTAGTACTTACTTTAACAAAAAATGATCTTGACAAAGCAAATAAAGACAA 7706 ATCACATAGACTTCCATTTTCTACTTTTTCTGAGGTTTCCTCTGGTCCTGGTATGAAGAA TGTATTTACCCAAAAGTGAAACATTTTGTCCTTTTTTAGCATCTTGTTCTGTTTGTGGAA 7707 GATGGGAGGACAAGTTCATGTATTTTGAGTTCCCTCAGCCGTTACCTGTGTGTGGTGAT ATCAAAGTAGAGTTCTTCCACAAACAGAACAAGATGCTAAAAAAGGACAAAATGTTTC ACTTTTGGGTA 7708 TTTGAAACTATTCCAATGTTCAGTGGCGGAACTTGCAATCCTCAGTTTGTGGTCTGCCA GCTAAAGGTGAAGATGTATTCCTCCAATTCAGGACCCACACGATGGGAGGACAAGTTC ATGTATTTTGAGTTCCCTCAGCCGTTACCTGTGTGTGGTGATATCAAAGTAGAGTTCT 7709 ACATCATCTTGTGAAACAACAGTGCCACTGGTCTATAATCCACATGATTCTTTACCAGG TAGCTATAGTAATACACATAGCGCCTCTGACTGGGAATAGTTACTCCCTTTTTGTCTCTG GTCCTTACTTCCCCATAGAAATCTAGGGCCTCTTGTGCCTTTAAAAATT 7710 AAGTGAAGATGACAATCATGTTGCAGCAATTCACTGTAAAGCTGGAAAGGGACGAACT GGTATAATGATTTATGCATATTTATTACATCGGGGCAA 7711 CGGTCCAGAGCCAAGCAGCGGCTGAGCGAGGGGCATCAGCTACCGCCAAGTCCAGAGC CATTTCCATCCTGCAGAGCCCCGCCACCAG 7712 CTGGTGGCGTAAGCAAAAGTGTCTTGACGATACAGCTAATTCAGAATCATTTTGTGGAC CAATATGATCCAACAATAGAGAATTCCTACAGGAAGCAAGTAGTAATTGATGGAGAAA 7713 CCCCCCCCCCCCCCCGCCCAGTCCTCATGTACTGGTCCTCATTGCATTGTACTCTTCTTG ACCTGTTGTGTCAAGAATATCCAAGAGACAGGT

REFERENCES

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SEQUENCES anaplastic lymphoma kinase (ALK) SEQ ID NO: 7714 1 mgaigllwllplllstaavgsgmgtgqragspaagpplqpreplsysrlqrkslavdfvv 61 pslfrvyardlllppssselkagrpeargslaldcapllrllgpapgvswtagspapaea 121 rtlsrvlkggsvrklrrakqlvlelgeeailegcvgppgeaavgllqfnlselfswwirq 181 gegrlrirlmpekkasevgregrlsaairasqprllfqifgtghsslesptnmpspspdy 241 ftwnltwimkdsfpflshrsryglecsfdfpceleyspplhdlrnqswswrripseeasq 301 mdlldgpgaerskemprgsflllntsadskhtilspwmrsssehctlavsvhrhlqpsgr 361 yiaqllphneaareillmptpgkhgwtvlqgrigrpdnpfrvaleyissgnrslsavdff 421 alkncsegtspgskmalqssftcwngtvlqlgqacdfhqdcaqgedesqmcrklpvgfyc 481 nfedgfcgwtqgtlsphtpqwqvrtlkdarfqdhqdhalllsttdvpasesatvtsatfp 541 apiksspcelrmswlirgvlrgnvslvlvenktgkeqgrmvwhvaayeglslwqwmvlpl 601 ldvsdrfwlqmvawwgqgsraivafdnisisldcyltisgedkilqntapksrnlfernp 661 nkelkpgensprqtpifdptvhwlfttcgasgphgptqaqcnnayqnsnlsvevgsegpl 721 kgiqiwkvpatdtysisgygaaggkggkntmmrshgvsvlgifnlekddmlyilvgqqge 781 dacpstnqliqkvcigennvieeeirvnrsvhewaggggggggatyvfkmkdgvpvplii 841 aaggggraygaktdtfhperlennssvlglngnsgaagggggwndntsllwagkslqega 901 tgghscpqamkkwgwetrggfggggggcssggggggyiggnaasnndpemdgedgvsfis 961 plgilytpalkvmeghgevnikhylncshcevdechmdpeshkvicfcdhgtvlaedgvs 1021 civsptpephlplslilsvvtsalvaalvlafsgimivyrrkhgelqamqmelqspeykl 1081 sklrtstimtdynpnycfagktssisdlkevprknitlirglghgafgevyegqvsgmpn 1141 dpsplqvavktlpevcseqdeldflmealiiskfnhqnivrcigvslqslprfillelma 1201 ggdlksflretrprpsqpsslamldllhvardiacgcqyleenhfihrdiaarnclltcp 1261 gpgrvakigdfgmardiyrasyyrkggcamlpvkwmppeafmegiftsktdtwsfgvllw 1321 eifslgympypsksnqevlefvtsggrmdppkncpgpvyrimtqcwqhqpedrpnfaiil 1381 erieyctqdpdvintalpieygplveeeekvpvrpkdpegvppllvsqqakreeerspaa 1441 ppplpttssgkaakkptaaeisvrvprgpavegghvnmafsqsnppselhkvhgsrnkpt 1501 slwnptygswftekptkknnpiakkephdrgnlglegsctvppnvatgrlpgasllleps 1561 sltanmkevplfrlrhfpcgnvnygyqqqglpleaatapgaghyedtilksknsmnqpgp ALK cDNA Sequence Reference SEQ ID NO: 7715 atgggagccatcgggctcctgtggctcctgccgctgctgctttccacggcagctgtgggc tccgggatggggaccggccagcgcgcgggctccccagctgcggggccgccgctgcagccc cgggagccactcagctactcgcgcctgcagaggaagagtctggcagttgacttcgtggtg ccctcgctcttccgtgtctacgcccgggacctactgctgccaccatcctcctcggagctg aaggctggcaggcccgaggcccgcggctcgctagctctggactgcgccccgctgctcagg ttgctggggccggcgccgggggtctcctggaccgccggttcaccagccccggcagaggcc cggacgctgtccagggtgctgaagggcggctccgtgcgcaagctccggcgtgccaagcag ttggtgctggagctgggcgaggaggcgatcttggagggttgcgtcgggccccccggggag gcggctgtggggctgctccagttcaatctcagcgagctgttcagttggtggattcgccaa ggcgaagggcgactgaggatccgcctgatgcccgagaagaaggcgtcggaagtgggcaga gagggaaggctgtccgcggcaattcgcgcctcccagccccgccttctcttccagatcttc gggactggtcatagctccttggaatcaccaacaaacatgccttctccttctcctgattat tttacatggaatctcacctggataatgaaagactccttccctttcctgtctcatcgcagc cgatatggtctggagtgcagctttgacttcccctgtgagctggagtattcccctccactg catgacctcaggaaccagagctggtcctggcgccgcatcccctccgaggaggcctcccag atggacttgctggatgggcctggggcagagcgttctaaggagatgcccagaggctccttt ctccttctcaacacctcagctgactccaagcacaccatcctgagtccgtggatgaggagc agcagtgagcactgcacactggccgtctcggtgcacaggcacctgcagccctctggaagg tacattgcccagctgctgccccacaacgaggctgcaagagagatcctcctgatgcccact ccagggaagcatggttggacagtgctccagggaagaatcgggcgtccagacaacccattt cgagtggccctggaatacatctccagtggaaaccgcagcttgtctgcagtggacttcttt gccctgaagaactgcagtgaaggaacatccccaggctccaagatggccctgcagagctcc ttcacttgttggaatgggacagtcctccagcttgggcaggcctgtgacttccaccaggac tgtgcccagggagaagatgagagccagatgtgccggaaactgcctgtgggtttttactgc aactttgaagatggcttctgtggctggacccaaggcacactgtcaccccacactcctcaa tggcaggtcaggaccctaaaggatgcccggttccaggaccaccaagaccatgctctattg ctcagtaccactgatgtccccgcttctgaaagtgctacagtgaccagtgctacgtttcct gcaccgatcaagagctctccatgtgagctccgaatgtcctggctcattcgtggagtcttg aggggaaacgtgtccttggtgctagtggagaacaaaaccgggaaggagcaaggcaggatg gtctggcatgtcgccgcctatgaaggcttgagcctgtggcagtggatggtgttgcctctc ctcgatgtgtctgacaggttctggctgcagatggtcgcatggtggggacaaggatccaga gccatcgtggcttttgacaatatctccatcagcctggactgctacctcaccattagcgga gaggacaagatcctgcagaatacagcacccaaatcaagaaacctgtttgagagaaaccca aacaaggagctgaaacccggggaaaattcaccaagacagacccccatctttgaccctaca gttcattggctgttcaccacatgtggggccagcgggccccatggccccacccaggcacag tgcaacaacgcctaccagaactccaacctgagcgtggaggtggggagcgagggccccctg aaaggcatccagatctggaaggtgccagccaccgacacctacagcatctcgggctacgga gctgctggcgggaaaggcgggaagaacaccatgatgcggtcccacggcgtgtctgtgctg ggcatcttcaacctggagaaggatgacatgctgtacatcctggttgggcagcagggagag gacgcctgccccagtacaaaccagttaatccagaaagtctgcattggagagaacaatgtg atagaagaagaaatccgtgtgaacagaagcgtgcatgagtgggcaggaggcggaggagga gggggtggagccacctacgtatttaagatgaaggatggagtgccggtgcccctgatcatt gcagccggaggtggtggcagggcctacggggccaagacagacacgttccacccagagaga ctggagaataactcctcggttctagggctaaacggcaattccggagccgcaggtggtgga ggtggctggaatgataacacttccttgctctgggccggaaaatctttgcaggagggtgcc accggaggacattcctgcccccaggccatgaagaagtgggggtgggagacaagagggggt ttcggagggggtggaggggggtgctcctcaggtggaggaggcggaggatatataggcggc aatgcagcctcaaacaatgaccccgaaatggatggggaagatggggtttccttcatcagt ccactgggcatcctgtacaccccagctttaaaagtgatggaaggccacggggaagtgaat attaagcattatctaaactgcagtcactgtgaggtagacgaatgtcacatggaccctgaa agccacaaggtcatctgcttctgtgaccacgggacggtgctggctgaggatggcgtctcc tgcattgtgtcacccaccccggagccacacctgccactctcgctgatcctctctgtggtg acctctgccctcgtggccgccctggtcctggctttctccggcatcatgattgtgtaccgc cggaagcaccaggagctgcaagccatgcagatggagctgcagagccctgagtacaagctg agcaagctccgcacctcgaccatcatgaccgactacaaccccaactactgctttgctggc aagacctcctccatcagtgacctgaaggaggtgccgcggaaaaacatcaccctcattcgg ggtctgggccatggcgcctttggggaggtgtatgaaggccaggtgtccggaatgcccaac gacccaagccccctgcaagtggctgtgaagacgctgcctgaagtgtgctctgaacaggac gaactggatttcctcatggaagccctgatcatcagcaaattcaaccaccagaacattgtt cgctgcattggggtgagcctgcaatccctgccccggttcatcctgctggagctcatggcg gggggagacctcaagtccttcctccgagagacccgccctcgcccgagccagccctcctcc ctggccatgctggaccttctgcacgtggctcgggacattgcctgtggctgtcagtatttg gaggaaaaccacttcatccaccgagacattgctgccagaaactgcctcttgacctgtcca ggccctggaagagtggccaagattggagacttcgggatggcccgagacatctacagggcg agctactatagaaagggaggctgtgccatgctgccagttaagtggatgcccccagaggcc ttcatggaaggaatattcacttctaaaacagacacatggtcctttggagtgctgctatgg gaaatcttttctcttggatatatgccataccccagcaaaagcaaccaggaagttctggag tttgtcaccagtggaggccggatggacccacccaagaactgccctgggcctgtataccgg ataatgactcagtgctggcaacatcagcctgaagacaggcccaactttgccatcattttg gagaggattgaatactgcacccaggacccggatgtaatcaacaccgctttgccgatagaa tatggtccacttgtggaagaggaagagaaagtgcctgtgaggcccaaggaccctgagggg gttcctcctctcctggtctctcaacaggcaaaacgggaggaggagcgcagcccagctgcc ccaccacctctgcctaccacctcctctggcaaggctgcaaagaaacccacagctgcagag atctctgttcgagtccctagagggccggccgtggaagggggacacgtgaatatggcattc tctcagtccaaccctccttcggagttgcacaaggtccacggatccagaaacaagcccacc agcttgtggaacccaacgtacggctcctggtttacagagaaacccaccaaaaagaataat cctatagcaaagaaggagccacacgacaggggtaacctggggctggagggaagctgtact gtcccacctaacgttgcaactgggagacttccgggggcctcactgctcctagagccctct tcgctgactgccaatatgaaggaggtacctctgttcaggctacgtcacttcccttgtggg aatgtcaattacggctaccagcaacagggcttgcccttagaagccgctactgcccctgga gctggtcattacgaggataccattctgaaaagcaagaatagcatgaaccagcctgggccc tga EGFR cDNA Sequence Reference SEQ ID NO: 7716 ATGCGACCCTCCGGGACGGCCGGGGCAGCGCTCCTGGCGCTGCTGGCTGCGCTCTGCCCG GCGAGTCGGCCTCTGGAGGAAAAGAAAGTTTGCCAAGGCACGAGTAACAACCTCACGCAG TTGGGCACTTTTGAAGATCATTTTCTCAGCCTCCAGAGGATGTTCAATAACTGTGAGGTG GTCCTTGGGAATTTGGAAATTACCTATGTGCAGAGGAATTATGATCTTTCCTTCTTAAAG ACCATCCAGGAGGTGGCTGGTTATGTCCTCATTGCCCTCAACACAGTGGAGCGAATTCCT TTGGAAAACCTGCAGATCATCAGAGGAAATATGTACTACGAAAATTCCTATGCCTTAGCA GTCTTATCTAACTATGATGCAAATAAAACCGGACTGAAGGAGCTGCCCATGAGAAATTTA CAGGAAATCCTGCATGGCGCCGTGCGGTTCAGCAACAACCCTGCCCTGTGCAACGTGGAG AGCATCCAGTGGCGGGACATAGTCAGCAGTGACTTTCTCAGCAACATGTCGATGGACTTC CAGAACCACCTGGGCAGCTGCCAAAAGTGTGATCCAAGCTGTCCCAATGGGAGCTGCTGG GGTGCAGGAGAGGAGAACTGCCAGAAACTGACCAAAATCATCTGTGCCCAGCAGTGCTCC GGGCGCTGCCGTGGCAAGTCCCCCAGTGACTGCTGCCACAACCAGTGTGCTGCAGGCTGC ACAGGCCCCCGGGAGAGCGACTGCCTGGTCTGCCGCAAATTCCGAGACGAAGCCACGTGC AAGGACACCTGCCCCCCACTCATGCTCTACAACCCCACCACGTACCAGATGGATGTGAAC CCCGAGGGCAAATACAGCTTTGGTGCCACCTGCGTGAAGAAGTGTCCCCGTAATTATGTG GTGACAGATCACGGCTCGTGCGTCCGAGCCTGTGGGGCCGACAGCTATGAGATGGAGGAA GACGGCGTCCGCAAGTGTAAGAAGTGCGAAGGGCCTTGCCGCAAAGTGTGTAACGGAATA GGTATTGGTGAATTTAAAGACTCACTCTCCATAAATGCTACGAATATTAAACACTTCAAA AACTGCACCTCCATCAGTGGCGATCTCCACATCCTGCCGGTGGCATTTAGGGGTGACTCC TTCACACATACTCCTCCTCTGGATCCACAGGAACTGGATATTCTGAAAACCGTAAAGGAA ATCACAGGGTTTTTGCTGATTCAGGCTTGGCCTGAAAACAGGACGGACCTCCATGCCTTT GAGAACCTAGAAATCATACGCGGCAGGACCAAGCAACATGGTCAGTTTTCTCTTGCAGTC GTCAGCCTGAACATAACATCCTTGGGATTACGCTCCCTCAAGGAGATAAGTGATGGAGAT GTGATAATTTCAGGAAACAAAAATTTGTGCTATGCAAATACAATAAACTGGAAAAAACTG TTTGGGACCTCCGGTCAGAAAACCAAAATTATAAGCAACAGAGGTGAAAACAGCTGCAAG GCCACAGGCCAGGTCTGCCATGCCTTGTGCTCCCCCGAGGGCTGCTGGGGCCCGGAGCCC AGGGACTGCGTCTCTTGCCGGAATGTCAGCCGAGGCAGGGAATGCGTGGACAAGTGCAAC CTTCTGGAGGGTGAGCCAAGGGAGTTTGTGGAGAACTCTGAGTGCATACAGTGCCACCCA GAGTGCCTGCCTCAGGCCATGAACATCACCTGCACAGGACGGGGACCAGACAACTGTATC CAGTGTGCCCACTACATTGACGGCCCCCACTGCGTCAAGACCTGCCCGGCAGGAGTCATG GGAGAAAACAACACCCTGGTCTGGAAGTACGCAGACGCCGGCCATGTGTGCCACCTGTGC CATCCAAACTGCACCTACGGATGCACTGGGCCAGGTCTTGAAGGCTGTCCAACGAATGGG CCTAAGATCCCGTCCATCGCCACTGGGATGGTGGGGGCCCTCCTCTTGCTGCTGGTGGTG GCCCTGGGGATCGGCCTCTTCATGCGAAGGCGCCACATCGTTCGGAAGCGCACGCTGCGG AGGCTGCTGCAGGAGAGGGAGCTTGTGGAGCCTCTTACACCCAGTGGAGAACCTCCCAAC CAAGCTCTCTTGAGGATCTTGAAGGAAACTGAATTCAAAAAGATCAAAGTGCTGGGCTCC GGTGCGTTCGGCACGGTGTATAAGGGACTCTGGATCCCAGAAGGTGAGAAAGTTAAAATT CCCGTCGCTATCAAGGAATTAAGAGAAGCAACATCTCCGAAAGCCAACAAGGAAATCCTC GATGAAGCCTACGTGATGGCCAGCGTGGACAACCCCCACGTGTGCCGCCTGCTGGGCATC TGCCTCACCTCCACCGTGCAGCTCATCACGCAGCTCATGCCCTTCGGCTGCCTCCTGGAC TATGTCCGGGAACACAAAGACAATATTGGCTCCCAGTACCTGCTCAACTGGTGTGTGCAG ATCGCAAAGGGCATGAACTACTTGGAGGACCGTCGCTTGGTGCACCGCGACCTGGCAGCC AGGAACGTACTGGTGAAAACACCGCAGCATGTCAAGATCACAGATTTTGGGCTGGCCAAA CTGCTGGGTGCGGAAGAGAAAGAATACCATGCAGAAGGAGGCAAAGTGCCTATCAAGTGG ATGGCATTGGAATCAATTTTACACAGAATCTATACCCACCAGAGTGATGTCTGGAGCTAC GGGGTGACTGTTTGGGAGTTGATGACCTTTGGATCCAAGCCATATGACGGAATCCCTGCC AGCGAGATCTCCTCCATCCTGGAGAAAGGAGAACGCCTCCCTCAGCCACCCATATGTACC ATCGATGTCTACATGATCATGGTCAAGTGCTGGATGATAGACGCAGATAGTCGCCCAAAG TTCCGTGAGTTGATCATCGAATTCTCCAAAATGGCCCGAGACCCCCAGCGCTACCTTGTC ATTCAGGGGGATGAAAGAATGCATTTGCCAAGTCCTACAGACTCCAACTTCTACCGTGCC CTGATGGATGAAGAAGACATGGACGACGTGGTGGATGCCGACGAGTACCTCATCCCACAG CAGGGCTTCTTCAGCAGCCCCTCCACGTCACGGACTCCCCTCCTGAGCTCTCTGAGTGCA ACCAGCAACAATTCCACCGTGGCTTGCATTGATAGAAATGGGCTGCAAAGCTGTCCCATC AAGGAAGACAGCTTCTTGCAGCGATACAGCTCAGACCCCACAGGCGCCTTGACTGAGGAC AGCATAGACGACACCTTCCTCCCAGTGCCTGAATACATAAACCAGTCCGTTCCCAAAAGG CCCGCTGGCTCTGTGCAGAATCCTGTCTATCACAATCAGCCTCTGAACCCCGCGCCCAGC AGAGACCCACACTACCAGGACCCCCACAGCACTGCAGTGGGCAACCCCGAGTATCTCAAC ACTGTCCAGCCCACCTGTGTCAACAGCACATTCGACAGCCCTGCCCACTGGGCCCAGAAA GGCAGCCACCAAATTAGCCTGGACAACCCTGACTACCAGCAGGACTTCTTTCCCAAGGAA GCCAAGCCAAATGGCATCTTTAAGGGCTCCACAGCTGAAAATGCAGAATACCTAAGGGTC GCGCCACAAAGCAGTGAATTTATTGGAGCATGA BRAF cDNA Sequence Reference SEQ ID NO: 7717 atggcggcgctgagcggtggcggtggtggcggcgcggagccgggccaggctctgttcaac ggggacatggagcccgaggccggcgccggcgccggcgccgcggcctcttcggctgcggac cctgccattccggaggaggtgtggaatatcaaacaaatgattaagttgacacaggaacat atagaggccctattggacaaatttggtggggagcataatccaccatcaatatatctggag gcctatgaagaatacaccagcaagctagatgcactccaacaaagagaacaacagttattg gaatctctggggaacggaactgatttttctgtttctagctctgcatcaatggataccgtt acatcttcttcctcttctagcctttcagtgctaccttcatctctttcagtttttcaaaat cccacagatgtggcacggagcaaccccaagtcaccacaaaaacctatcgttagagtcttc ctgcccaacaaacagaggacagtggtacctgcaaggtgtggagttacagtccgagacagt ctaaagaaagcactgatgatgagaggtctaatcccagagtgctgtgctgtttacagaatt caggatggagagaagaaaccaattggttgggacactgatatttcctggcttactggagaa gaattgcatgtggaagtgttggagaatgttccacttacaacacacaactttgtacgaaaa acgtttttcaccttagcattttgtgacttttgtcgaaagctgcttttccagggtttccgc tgtcaaacatgtggttataaatttcaccagcgttgtagtacagaagttccactgatgtgt gttaattatgaccaacttgatttgctgtttgtctccaagttctttgaacaccacccaata ccacaggaagaggcgtccttagcagagactgccctaacatctggatcatccccttccgca cccgcctcggactctattgggccccaaattctcaccagtccgtctccttcaaaatccatt ccaattccacagcccttccgaccagcagatgaagatcatcgaaatcaatttgggcaacga gaccgatcctcatcagctcccaatgtgcatataaacacaatagaacctgtcaatattgat gacttgattagagaccaaggatttcgtggtgatggaggatcaaccacaggtttgtctgct accccccctgcctcattacctggctcactaactaacgtgaaagccttacagaaatctcca ggacctcagcgagaaaggaagtcatcttcatcctcagaagacaggaatcgaatgaaaaca cttggtagacgggactcgagtgatgattgggagattcctgatgggcagattacagtggga caaagaattggatctggatcatttggaacagtctacaagggaaagtggcatggtgatgtg gcagtgaaaatgttgaatgtgacagcacctacacctcagcagttacaagccttcaaaaat gaagtaggagtactcaggaaaacacgacatgtgaatatcctactcttcatgggctattcc acaaagccacaactggctattgttacccagtggtgtgagggctccagcttgtatcaccat ctccatatcattgagaccaaatttgagatgatcaaacttatagatattgcacgacagact gcacagggcatggattacttacacgccaagtcaatcatccacagagacctcaagagtaat aatatatttcttcatgaagacctcacagtaaaaataggtgattttggtctagctacagtg aaatctcgatggagtgggtcccatcagtttgaacagttgtctggatccattttgtggatg gcaccagaagtcatcagaatgcaagataaaaatccatacagctttcagtcagatgtatat gcatttggaattgttctgtatgaattgatgactggacagttaccttattcaaacatcaac aacagggaccagataatttttatggtgggacgaggatacctgtctccagatctcagtaag gtacggagtaactgtccaaaagccatgaagagattaatggcagagtgcctcaaaaagaaa agagatgagagaccactctttccccaaattctcgcctctattgagctgctggcccgctca ttgccaaaaattcaccgcagtgcatcagaaccctccttgaatcgggctggtttccaaaca gaggattttagtctatatgcttgtgcttctccaaaaacacccatccaggcagggggatat ggtgcgtttcctgtccactga KRAS cDNA Sequence Reference SEQ ID NO: 7718 atgactgaatataaacttgtggtagttggagctggtggcgtaggcaagagtgccttgacg atacagctaattcagaatcattttgtggacgaatatgatccaacaatagaggattcctac aggaagcaagtagtaattgatggagaaacctgtctcttggatattctcgacacagcaggt caagaggagtacagtgcaatgagggaccagtacatgaggactggggagggctttctttgt gtatttgccataaataatactaaatcatttgaagatattcaccattatagagaacaaatt aaaagagttaaggactctgaagatgtacctatggtcctagtaggaaataaatgtgatttg ccttctagaacagtagacacaaaacaggctcaggacttagcaagaagttatggaattcct tttattgaaacatcagcaaagacaagacagggtgttgatgatgccttctatacattagtt cgagaaattcgaaaacataaagaaaagatgagcaaagatggtaaaaagaagaaaaagaag tcaaagacaaagtgtgtaattatgtaa 

What is claimed is:
 1. A kinase inhibitor resistance panel comprising one or more primer sets from one or more of the genes selected from group of genes comprising KRAS, BRAF, EGFR, ALK, and KIT.
 2. The kinase inhibitor resistance panel of claim 1, wherein at least one primer sets hybridizes and amplifies nucleic acid from exon 1 or 2 of KRAS.
 3. The kinase inhibitor panel of claim 2, wherein one or more KRAS hybridizing primers or primer sets comprise one or more of the primers of Tables 10 and/or
 14. 4. The kinase inhibitor panel of claim 2, wherein one or more KRAS hybridizing primers or primer sets comprise one or more of the primers of SEQ ID NOs: 4601-5200 and 7181-7610.
 5. The kinase inhibitor resistance panel of claim 1, wherein at least one primer sets hybridizes and amplifies nucleic acid from exon 18, 19, 20, 21 or 22 of EGFR.
 6. The kinase inhibitor panel of claim 5, wherein one or more EGFR hybridizing primers or primer sets comprise one or more of the primers of Tables 8 and/or
 12. 7. The kinase inhibitor panel of claim 6, wherein one or more EGFR hybridizing primers or primer sets comprise one or more of the primers of SEQ ID NOs: 1641-2440 and 5819-6524.
 8. The kinase inhibitor resistance panel of claim 1, wherein at least one primer sets hybridizes and amplifies nucleic acid from exon 21, 22, 23, 24, or 25 of ALK.
 9. The kinase inhibitor panel of claim 8, wherein one or more ALK hybridizing primers or primer sets comprise one or more of the primers of Tables 7 and/or
 11. 10. The kinase inhibitor panel of claim 9, wherein one or more ALK hybridizing primers or primer sets comprise one or more of the primers of SEQ ID NOs: 1-1640 and 5201-5818.
 11. The kinase inhibitor resistance panel of claim 1, wherein at least one primer sets hybridizes and amplifies nucleic acid from exon 8, 9, 10, 11, 12, 13, or 17 of KIT.
 12. The kinase inhibitor panel of claim 11, wherein one or more KIT hybridizing primers or primer sets comprise one or more of the primers of Table
 9. 13. The kinase inhibitor panel of claim 12, wherein one or more KIT hybridizing primers or primer sets comprise one or more of the primers of SEQ ID NOs: 2441-4600.
 14. The kinase inhibitor resistance panel of claim 1, wherein at least one primer sets hybridizes and amplifies nucleic acid from exon 10, 11, 13, 14, or 15 of BRAF.
 15. The kinase inhibitor panel of claim 14, wherein one or more BRAF hybridizing primers or primer sets comprise one or more of the primers of Table
 13. 16. The kinase inhibitor panel of claim 15, wherein one or more BRAF hybridizing primers or primer sets comprise one or more of the primers of SEQ ID NOs: 6525-7180.
 17. The kinase inhibitor panel of claim 1, wherein the panel comprises two or more primer sets for one or more of the genes selected from group of genes comprising KRAS, BRAF, EGFR, ALK, and KIT.
 18. The kinase inhibitor panel of claim 1, wherein the panel comprises 3, 4, 5, 6, 7, 8, 9, 10, or more primer sets for one or more of the genes selected from group of genes comprising KRAS, BRAF, EGFR, ALK, and KIT.
 19. The kinase inhibitor panel of claim 1, wherein the panel comprises one or more primer sets for 2, 3, 4, of all 5 of the genes selected from group of genes comprising KRAS, BRAF, EGFR, ALK, and KIT.
 20. The kinase inhibitor of claim 1, wherein the kinase inhibitor resistance is resistance to an ALK kinase inhibitor.
 21. The kinase inhibitor resistance panel of claim 20, wherein the kinase inhibitor is selected from the group consisting of crizotinib, afatinib, Axitinib, bevacizumab, Bosutinib, Cetuximab, Dasatinib, Erlotinib, Fostamati nib, Gefitinib, Imatinib, Lapatinib, Lenvatinib, Nilotinib, Panitumumab, Pazopanib, Pegaptanib, Ranibizumab, Ruxolitinib, Sorafenib, Sunitinib, Trastuzumab, and Vemurafenib.
 22. The kinase inhibitor resistance panel of claim 21, wherein the kinase inhibitor comprises crizotinib.
 23. The kinase inhibitor panel of claim 1, wherein the one or more primer sets amplify one or more mutations associated with kinase inhibitor resistance, wherein the mutation is identified in Table 2, 3, 4, 5, or
 6. 24. A method for the detection of kinase inhibitor resistance comprising obtaining a tissue sample from a subject with a cancer and conducting a high throughput sequencing (also known as next generation sequencing) reaction on the sample using the kinase inhibitor resistant panel of claim 1, wherein the presence of a mutation in the nucleic acid sequence of a gene associated with kinase inhibitor resistance indicates that that the cancer is resistant or will become resistant to a kinase inhibitor.
 25. A method for the detection of kinase inhibitor resistance comprising obtaining a tissue sample from a subject with a cancer and conducting a high throughput sequencing (also known as next generation sequencing) reaction on the sample using one or more primer sets or primer panels with primer sets that specifically hybridizes to one or more of the genes selected from the group consisting ofALK, KRAS, EGFR, KIT, and BRAF, wherein the presence of a mutation in the nucleic acid sequence of a gene associated with kinase inhibitor resistance indicates that that the cancer is resistant or will become resistant to a kinase inhibitor.
 26. The method of claim 25, wherein at least one primer sets hybridizes and amplifies nucleic acid from exon 1 or 2 of KRAS.
 27. The method of claim 26, wherein one or more KRAS hybridizing primers or primer sets comprise one or more of the primers of Tables 10 and/or
 14. 28. The method of claim 26, wherein one or more KRAS hybridizing primers or primer sets comprise one or more of the primers of SEQ ID NOs: 4601-5200 and 7181-7610.
 29. The method of claim 25, wherein at least one primer sets hybridizes and amplifies nucleic acid from exon 18, 19, 20, 21 or 22 of EGFR.
 30. The method of claim 29, wherein one or more EGFR hybridizing primers or primer sets comprise one or more of the primers of Tables 8 and/or
 12. 31. The method of claim 29, wherein one or more EGFR hybridizing primers or primer sets comprise one or more of the primers of SEQ ID NOs: 1641-2440 and 5819-6524.
 32. The method of claim 25, wherein at least one primer sets hybridizes and amplifies nucleic acid from exon 21, 22, 23, 24, or 25 of ALK.
 33. The method of claim 32, wherein one or more ALK hybridizing primers or primer sets comprise one or more of the primers of Tables 7 and/or
 11. 34. The method of claim 32, wherein one or more ALK hybridizing primers or primer sets comprise one or more of the primers of SEQ ID NOs: 1-1640 and 5201-5818.
 35. The method of claim 25, wherein at least one primer sets hybridizes and amplifies nucleic acid from exon 8, 9, 10, 11, 12, 13, or 17 of KIT.
 36. The method of claim 35, wherein one or more KIT hybridizing primers or primer sets comprise one or more of the primers of Table
 9. 37. The method of claim 35, wherein one or more KIT hybridizing primers or primer sets comprise one or more of the primers of SEQ ID NOs: 2441-4600.
 38. The method of claim 25, wherein at least one primer sets hybridizes and amplifies nucleic acid from exon 10, 11, 13, 14, or 15 of BRAF.
 39. The method of claim 38, wherein one or more BRAF hybridizing primers or primer sets comprise one or more of the primers of Table
 13. 40. The method of claim 38, wherein one or more BRAF hybridizing primers or primer sets comprise one or more of the primers of SEQ ID NOs: 6525-7180.
 41. The method of claim 25, wherein the panel comprises two or more primer sets for one or more of the genes selected from group of genes comprising KRAS, BRAF, EGFR, ALK, and KIT.
 42. The method of claim 25, wherein the panel comprises 3, 4, 5, 6, 7, 8, 9, 10, or more primer sets for one or more of the genes selected from group of genes comprising KRAS, BRAF, EGFR, ALK, and KIT.
 43. The method of claim 25, wherein the panel comprises one or more primer sets for 2, 3, 4, of all 5 of the genes selected from group of genes comprising KRAS, BRAF, EGFR, ALK, and KIT.
 44. The method of claim 25, wherein the kinase inhibitor resistance is resistance to an ALK kinase inhibitor.
 45. The method of claim 25, wherein the kinase inhibitor is selected from the group consisting of crizotinib, afatinib, Axitinib, bevacizumab, Bosutinib, Cetuximab, Dasatinib, Erlotinib, Fostamati nib, Gefitinib, Imatinib, Lapatinib, Lenvatinib, Nilotinib, Panitumumab, Pazopanib, Pegaptanib, Ranibizumab, Ruxolitinib, Sorafenib, Sunitinib, Trastuzumab, and Vemurafenib.
 46. The method of claim 45, wherein the kinase inhibitor comprises crizotinib.
 47. The method of claim 25, wherein the one or more primer sets amplify one or more mutations associated with kinase inhibitor resistance, wherein the mutation is identified in Table 2, 3, 4, 5, or
 6. 